菲律宾蛤仔（Venerupis philippinarum）在苯并（a）芘胁迫下差异基因的筛选与分子生物标志物的研究
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摘要
本文根据目前我国近海多环芳烃污染现状，以近海养殖重要经济贝类菲律宾蛤仔（Venerupis philippinarum）为研究对象，克隆了菲律宾蛤仔芳烃受体介导的解毒代谢途径几种关键基因，包括芳烃受体蛋白（AhR）、芳烃受体核转位因子（ARNT）、热休克蛋白90（HSP90）。研究了毒性最强的多环芳烃（PAHs）——苯并（a）芘（BaP）对菲律宾蛤仔组织解毒代谢相关基因的表达调控规律，通过对在BaP胁迫下菲律宾蛤仔解毒代谢相关基因的克隆及基因表达分析，探究双壳贝类PAHs解毒代谢机制；构建了菲律宾蛤仔PAHs消减cDNA文库，筛选、鉴定了差异表达基因，结合室内实验和海区现场验证，初步建立了基于菲律宾蛤仔的PAHs分子生物标志物监测技术，为海洋环境污染监测和水产品安全评价提供基础数据和技术支持。
1菲律宾蛤仔芳烃受体（AhR）介导的关键基因克隆与表达分析
菲律宾蛤仔芳烃受体蛋白（VpAhR）的cDNA全序列长2870bp，开放阅读框（ORF）长为2391bp，共编码796个氨基酸残基，不具备信号肽，属于胞内蛋白，含有2个核定位序列（NLS）：K45RRRR49和P67SKRHRE73。BLAST分析表明VpAhR与其它物种AhR基因具有较高的同源性。VpAhR含有1个碱性螺旋-环-螺旋结构（bHLH domain）和2个配体结合区（PAS domain），属于bHLH-PAS转录因子家族。序列分析显示，VpAhR的配体结合区（a.a.220–383）与砂海螂（M. arenaria）和斑马纹贻贝（D. polymorpha）AhR的配体结合区相似性很高。实时定量PCR分析表明VpAhR在菲律宾蛤仔的鳃、消化盲囊、外套膜和闭壳肌中均有表达，鳃组织中表达量最高。
菲律宾蛤仔ARNT片段长485bp，编码161个氨基酸残基，该序列含一个由44个氨基酸构成的PAS结构域（a.a.7-50），该区域与其他物种ARNT的PAS结构域相似性较高；VpARNT在菲律宾蛤仔消化盲囊中表达最高，在鳃、外套膜和闭壳肌中也均有分布。HSP90是分子伴侣家族的重要一员，菲律宾蛤仔的HSP90基因片段cDNA长687bp，编码299个氨基酸，具有HSP90家族保守信号区（GVVDSEDLPLNISRE），与其他物种HSP90氨基酸序列具有很高相似性，与栉孔扇贝（C. farreri）和海湾扇贝（A. irradians）的相似性均为99%，菲律宾蛤仔HSP90在鳃组织中表达量最高；不同浓度BaP（0.01、0.2μg/L）染毒处理3d、10d，菲律宾蛤仔鳃组织中AhR、ARNT、HSP90和谷胱甘肽硫转移酶（GSTpi）的基因表达水平随染毒浓度增大和处理时间延长而逐渐升高；除10d时0.2μg/L组细胞色素P4504（CYP4）基因表达被极显著（P＜0.01）上调外，其余处理组CYP4mRNA表达水平在染毒期间与对照组差异不显著（P＞0.05）。实验结果表明，BaP与AhR介导的菲律宾蛤仔解毒代谢相关基因AhR、ARNT、HSP90和GSTpi的mRNA表达具有明显的剂量—效应关系，符合生物标志物筛选原则，适于进行进一步的分析和现场验证。
2菲律宾蛤仔在苯并（a）芘胁迫下cDNA消减文库的构建与分析
利用抑制性消减杂交技术（SSH）构建了BaP（1μg/L）胁迫下菲律宾蛤仔正、反向cDNA消减文库，分别获得194个和93个重组克隆，测序分析显示，正、反向文库分别得到168个和83个EST序列，经同源序列相似性比对，正向cDNA文库中有31种基因与已知功能基因具有较高同源性，12种为假定蛋白，2种为未知蛋白；反向文库中已知功能基因10种，假定蛋白4种。所有ESTs序列信息均已提交Genbank数据库，cDNA消减文库编号为LIBEST_027169。进一步序列比对、功能注释及分类显示，通过SSH筛选到的菲律宾蛤仔差异表达基因主要为药物代谢、杂环化合物代谢、细胞分解代谢、抗生素响应等代谢过程相关基因。
3菲律宾蛤仔在苯并（a）芘胁迫下分子生物标志物的筛选与验证
设置一系列BaP染毒浓度（0.25、1和4μg/L），利用实时定量PCR检测染毒处理10d时菲律宾蛤仔解毒代谢相关基因、SSH-selected差异表达基因及几种其他功能基因的mRNA表达。结果显示，测定的28种基因中Defensin、Serine protease、Thioesterprotein、GST C、Mn-SOD、ARNT等12种基因被BaP诱导表达上调，且与染毒浓度呈线性正相关关系，因此适于作为潜在的分子生物标志物进行进一步研究与验证。
在胶州湾和胶南附近海域各选择一个实验站点（S1和S2），于2011年12月采集各站点表层海水及菲律宾蛤仔样品。高效液相色谱法（HPLC）测定海水中PAHs含量和菲律宾蛤仔软体部中PAHs的累积量。结果显示，S1站点海水和菲律宾蛤仔组织中的总PAHs和BaP含量均高于S2站点；实时定量PCR分析表明，除HSP90和AhR外，PAHs含量较高的S1站点的其余10种分子生物标志物基因表达水平明显高于S2站点，表现出一定的剂量—效应关系。由此表明，菲律宾蛤仔12种基因基本符合生物标志物筛选原则，适于作为分子生物标志物用于海洋环境PAHs污染监测及毒性效应评估。
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminantsand the environmental PAHs concentrations have been increasing particularly in aquaticenvironment. In present study, the toxic effect of benzo(a)pyrene (BaP) on clam Venerupisphilippinarum was studied and the molecular biomarkers were selected for PAHs pollutionassessment. The key genes of aryl hydrocarbon receptor (AhR)-mediated detoxificationmechanism, including AhR, aryl hydrocarbon receptor nuclear translocator (ARNT) and heatshock protein90(HSP90) were cloned and the effects of BaP on gene expression was studied inorder to explore the mechanism of detoxification of the clam. A subtracted cDNA library ofVenerupis philippinarum exposed to BaP was constructed by suppression subtractivehybridisation (SSH) for identification of differential expression genes. The molecular biomarkersof clam were selected and confirmed by analysis of gene expression. The resulting data mayprovide basic genomic information of the bivalve and the useful technology of biomonitoring ofPAHs by bivalves was supplied for PAHs pollution in aquatic environment.
1Cloning and expression of the key genes of aryl hydrocarbon receptor (AhR)-mediateddetoxication mechanism of Venerupis philippinarum exposed to benzo(a)pyrene
The full-length VpAhR cDNA is2870bp, containing an open reading frame (ORF) of2391bp which encoded a polypeptide of796amino acids. The predicted amino acidsequences contain regions characteristic of AhRs from other species including basichelix-loop-helix (bHLH) and Per-ARNT-Sim (PAS) domains. Two nuclear translocationsequences can be identifed in the VpAhR sequence. Blast analysis showed that the deducedamino acid sequence of VpAhR shared high identity with other AhRs. The clam AhR mRNAexpression was detected in all the adult tissues tested (gill, digestive gland, adductor muscle andmantle) and highest transcription level was observed in gill compared to other tissues.
The partial length VpARNT cDNA is485bp, containing an ORF of2391bp. The ORF encode a peptide of161amino acids. The predicted amino acid sequences contain a PAS domainwhich has a high identity with other species’ PAS domains. VpARNT mRNA expression wasdetected in all the adult tissues and the highest transcription level was observed in digestive gland.HSP90was an important member of molecular chaperone family. The cDNA partial length ofVpHSP90is687bp which is coding a peptide of299amino acids. An HSP90family conservedsignal region (GVVDSEDLPLNISRE) was detected in VpHSP90and it has a high identity withother species’ HSP90, such as C. farreri and A. irradians. The highest mRNA expression ofVpHSP90was detected in gill.
Clams were exposed to different concentrations of BaP (0.01and0.2μg/L) for10d. Thegene expression of AhR, ARNT, HSP90and GST were increased with the enhancement ofBaP concentration and prolong of the experimental period, while there was no significantdifference of CYP4gene expression between control and BaP treatments. The potentialmolecular biomarkers were selected for further study.
2Construction and analysis of subtractive cDNA library of Venerupis philippinarumexposed to benzo(a)pyrene
Forward and reverse subtracted cDNA library of Venerupis philippinarum exposed to BaPwere constructed. One hundred and ninety-four and ninety-three recombinants were collectedseparately from forward and reverse subtracted cDNA library. Thirty-one unigenes and twelvehypothetical proteins were identified from forward cDNA library and there were ten unigenesand four hypothetical proteins in reverse cDNA library. The cDNA library accession number ofthe clam is LIBEST_027169. Analysis of multiple alignment and annotation showed that most ofthe SSH-selected genes were related to drug metabolic process, heterocycle metabolic process,cellular catabolic process, amine metabolic process, cellular nitrogen compound metabolicprocess and response to antibiotic.
3Screening and confirmation of molecular biomarkers of Venerupis philippinarumexposed to benzo(a)pyrene
A BaP (0.025,1and4μg/L) exposure for10d was conducted and the gene expression ofAhR related genes, SSH-selected genes and some functional genes of clam were tested by realtime quantitative PCR. Results showed that mRNA expression of twelve genes includingDefensin、Serine protease、Thioester protein、GST C、Mn-SOD、ARNT et al., wereup-regulated by BaP exposure and there was a significant linear relationship between BaP concentration and the gene expression, which indicated that these genes were suitablemolecular biomarkers of clam for PAHs assessment.
Two sampling sites were selected according to the different PAHs pollution situation of thesea areas in Jiaozhou Bay and Dragon Bay. The PAHs contents of surface seawater and tissue ofclam and gene expression of biomarkers of clam sampled from different sites were analyzed.Results showed that the contents of total PAHs and BaP were higher in S1than S2. The geneexpression level of clams collected from S1was higher than S2which is similar to the results oflaboratory expreiment. It is indicated that the twelve genes were suitable molecular biomarkersfor PAHs pollution assessment.
1菲律宾蛤仔芳烃受体（AhR）介导的关键基因克隆与表达分析
菲律宾蛤仔芳烃受体蛋白（VpAhR）的cDNA全序列长2870bp，开放阅读框（ORF）长为2391bp，共编码796个氨基酸残基，不具备信号肽，属于胞内蛋白，含有2个核定位序列（NLS）：K45RRRR49和P67SKRHRE73。BLAST分析表明VpAhR与其它物种AhR基因具有较高的同源性。VpAhR含有1个碱性螺旋-环-螺旋结构（bHLH domain）和2个配体结合区（PAS domain），属于bHLH-PAS转录因子家族。序列分析显示，VpAhR的配体结合区（a.a.220–383）与砂海螂（M. arenaria）和斑马纹贻贝（D. polymorpha）AhR的配体结合区相似性很高。实时定量PCR分析表明VpAhR在菲律宾蛤仔的鳃、消化盲囊、外套膜和闭壳肌中均有表达，鳃组织中表达量最高。
菲律宾蛤仔ARNT片段长485bp，编码161个氨基酸残基，该序列含一个由44个氨基酸构成的PAS结构域（a.a.7-50），该区域与其他物种ARNT的PAS结构域相似性较高；VpARNT在菲律宾蛤仔消化盲囊中表达最高，在鳃、外套膜和闭壳肌中也均有分布。HSP90是分子伴侣家族的重要一员，菲律宾蛤仔的HSP90基因片段cDNA长687bp，编码299个氨基酸，具有HSP90家族保守信号区（GVVDSEDLPLNISRE），与其他物种HSP90氨基酸序列具有很高相似性，与栉孔扇贝（C. farreri）和海湾扇贝（A. irradians）的相似性均为99%，菲律宾蛤仔HSP90在鳃组织中表达量最高；不同浓度BaP（0.01、0.2μg/L）染毒处理3d、10d，菲律宾蛤仔鳃组织中AhR、ARNT、HSP90和谷胱甘肽硫转移酶（GSTpi）的基因表达水平随染毒浓度增大和处理时间延长而逐渐升高；除10d时0.2μg/L组细胞色素P4504（CYP4）基因表达被极显著（P＜0.01）上调外，其余处理组CYP4mRNA表达水平在染毒期间与对照组差异不显著（P＞0.05）。实验结果表明，BaP与AhR介导的菲律宾蛤仔解毒代谢相关基因AhR、ARNT、HSP90和GSTpi的mRNA表达具有明显的剂量—效应关系，符合生物标志物筛选原则，适于进行进一步的分析和现场验证。
2菲律宾蛤仔在苯并（a）芘胁迫下cDNA消减文库的构建与分析
利用抑制性消减杂交技术（SSH）构建了BaP（1μg/L）胁迫下菲律宾蛤仔正、反向cDNA消减文库，分别获得194个和93个重组克隆，测序分析显示，正、反向文库分别得到168个和83个EST序列，经同源序列相似性比对，正向cDNA文库中有31种基因与已知功能基因具有较高同源性，12种为假定蛋白，2种为未知蛋白；反向文库中已知功能基因10种，假定蛋白4种。所有ESTs序列信息均已提交Genbank数据库，cDNA消减文库编号为LIBEST_027169。进一步序列比对、功能注释及分类显示，通过SSH筛选到的菲律宾蛤仔差异表达基因主要为药物代谢、杂环化合物代谢、细胞分解代谢、抗生素响应等代谢过程相关基因。
3菲律宾蛤仔在苯并（a）芘胁迫下分子生物标志物的筛选与验证
设置一系列BaP染毒浓度（0.25、1和4μg/L），利用实时定量PCR检测染毒处理10d时菲律宾蛤仔解毒代谢相关基因、SSH-selected差异表达基因及几种其他功能基因的mRNA表达。结果显示，测定的28种基因中Defensin、Serine protease、Thioesterprotein、GST C、Mn-SOD、ARNT等12种基因被BaP诱导表达上调，且与染毒浓度呈线性正相关关系，因此适于作为潜在的分子生物标志物进行进一步研究与验证。
在胶州湾和胶南附近海域各选择一个实验站点（S1和S2），于2011年12月采集各站点表层海水及菲律宾蛤仔样品。高效液相色谱法（HPLC）测定海水中PAHs含量和菲律宾蛤仔软体部中PAHs的累积量。结果显示，S1站点海水和菲律宾蛤仔组织中的总PAHs和BaP含量均高于S2站点；实时定量PCR分析表明，除HSP90和AhR外，PAHs含量较高的S1站点的其余10种分子生物标志物基因表达水平明显高于S2站点，表现出一定的剂量—效应关系。由此表明，菲律宾蛤仔12种基因基本符合生物标志物筛选原则，适于作为分子生物标志物用于海洋环境PAHs污染监测及毒性效应评估。
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminantsand the environmental PAHs concentrations have been increasing particularly in aquaticenvironment. In present study, the toxic effect of benzo(a)pyrene (BaP) on clam Venerupisphilippinarum was studied and the molecular biomarkers were selected for PAHs pollutionassessment. The key genes of aryl hydrocarbon receptor (AhR)-mediated detoxificationmechanism, including AhR, aryl hydrocarbon receptor nuclear translocator (ARNT) and heatshock protein90(HSP90) were cloned and the effects of BaP on gene expression was studied inorder to explore the mechanism of detoxification of the clam. A subtracted cDNA library ofVenerupis philippinarum exposed to BaP was constructed by suppression subtractivehybridisation (SSH) for identification of differential expression genes. The molecular biomarkersof clam were selected and confirmed by analysis of gene expression. The resulting data mayprovide basic genomic information of the bivalve and the useful technology of biomonitoring ofPAHs by bivalves was supplied for PAHs pollution in aquatic environment.
1Cloning and expression of the key genes of aryl hydrocarbon receptor (AhR)-mediateddetoxication mechanism of Venerupis philippinarum exposed to benzo(a)pyrene
The full-length VpAhR cDNA is2870bp, containing an open reading frame (ORF) of2391bp which encoded a polypeptide of796amino acids. The predicted amino acidsequences contain regions characteristic of AhRs from other species including basichelix-loop-helix (bHLH) and Per-ARNT-Sim (PAS) domains. Two nuclear translocationsequences can be identifed in the VpAhR sequence. Blast analysis showed that the deducedamino acid sequence of VpAhR shared high identity with other AhRs. The clam AhR mRNAexpression was detected in all the adult tissues tested (gill, digestive gland, adductor muscle andmantle) and highest transcription level was observed in gill compared to other tissues.
The partial length VpARNT cDNA is485bp, containing an ORF of2391bp. The ORF encode a peptide of161amino acids. The predicted amino acid sequences contain a PAS domainwhich has a high identity with other species’ PAS domains. VpARNT mRNA expression wasdetected in all the adult tissues and the highest transcription level was observed in digestive gland.HSP90was an important member of molecular chaperone family. The cDNA partial length ofVpHSP90is687bp which is coding a peptide of299amino acids. An HSP90family conservedsignal region (GVVDSEDLPLNISRE) was detected in VpHSP90and it has a high identity withother species’ HSP90, such as C. farreri and A. irradians. The highest mRNA expression ofVpHSP90was detected in gill.
Clams were exposed to different concentrations of BaP (0.01and0.2μg/L) for10d. Thegene expression of AhR, ARNT, HSP90and GST were increased with the enhancement ofBaP concentration and prolong of the experimental period, while there was no significantdifference of CYP4gene expression between control and BaP treatments. The potentialmolecular biomarkers were selected for further study.
2Construction and analysis of subtractive cDNA library of Venerupis philippinarumexposed to benzo(a)pyrene
Forward and reverse subtracted cDNA library of Venerupis philippinarum exposed to BaPwere constructed. One hundred and ninety-four and ninety-three recombinants were collectedseparately from forward and reverse subtracted cDNA library. Thirty-one unigenes and twelvehypothetical proteins were identified from forward cDNA library and there were ten unigenesand four hypothetical proteins in reverse cDNA library. The cDNA library accession number ofthe clam is LIBEST_027169. Analysis of multiple alignment and annotation showed that most ofthe SSH-selected genes were related to drug metabolic process, heterocycle metabolic process,cellular catabolic process, amine metabolic process, cellular nitrogen compound metabolicprocess and response to antibiotic.
3Screening and confirmation of molecular biomarkers of Venerupis philippinarumexposed to benzo(a)pyrene
A BaP (0.025,1and4μg/L) exposure for10d was conducted and the gene expression ofAhR related genes, SSH-selected genes and some functional genes of clam were tested by realtime quantitative PCR. Results showed that mRNA expression of twelve genes includingDefensin、Serine protease、Thioester protein、GST C、Mn-SOD、ARNT et al., wereup-regulated by BaP exposure and there was a significant linear relationship between BaP concentration and the gene expression, which indicated that these genes were suitablemolecular biomarkers of clam for PAHs assessment.
Two sampling sites were selected according to the different PAHs pollution situation of thesea areas in Jiaozhou Bay and Dragon Bay. The PAHs contents of surface seawater and tissue ofclam and gene expression of biomarkers of clam sampled from different sites were analyzed.Results showed that the contents of total PAHs and BaP were higher in S1than S2. The geneexpression level of clams collected from S1was higher than S2which is similar to the results oflaboratory expreiment. It is indicated that the twelve genes were suitable molecular biomarkersfor PAHs pollution assessment.
引文
[1] Akbay C V, Shamsi S A, Warner I M. Separation of Monomethyl-benz[a]anthracene isomers using Cyclodextrin-Modified Electrokinetic Chromatography.Journal of Chromatography A,2001,910(1):147~155
[2] Altschul S F, Madden T L, Sch ffer A A, et al. Gapped BLAST and PSI-BLAST: anew generation of protein database search programs. Nucleic Acids Research,1997,25(17):3389~3402
[3] Atchley W, Fitch W M. A natural classification of the basic helix–loop–helix classof transcription factors. Evolution,1997,94:5172~5176
[4] Berg P, Pongratz I. Differential Usage of nuclear export sequences regulatesintracellular localization of the Dioxin (Aryl Hydrocarbon) receptor. Journal ofBiological Chemistry,2001,276(46):43231~43238
[5] Bollérot K, Angelier N, Coumailleau P. Molecular cloning and embryonicexpression of the Xenopus Arnt gene. Mechanisms of Development,2001,108:227~231
[6] Booij K, Hoedemaker J R, Bakker J F. Dissolved PCEs, PAHs, and HCB in porewaters and overlying waters of contaminated harbor sediments. EnvironmentalScience&Technology,2003,37(18):4213~4220
[7] Boulanger N, Lowenberger C, Volf P, et al. Characterization of a defensin from thesand fly Phlebotomus duboscqi induced by challenge with bacteria or theprotozoan parasite Leishmania major. Infection and Immunity,2004,72:7140~7146
[8] Brown D J, Beneden R J V, Clark G C. Identification of two binding proteins forhalogenated aromatic hydrocarbons in the hard-shell clam, Mercenariamercenaria. Archives of Biochemistry and Biophysics,1995,319(1):217~224
[9] Brown M, Davies I M, Moffat C F, et al. Application of SSH and a macroarrayto investigate altered gene expression in Mytilus edulis in response to exposure tobenzo[a]pyrene. Marine Environmental Research,2006,62: S128~S135
[10] Bulet P, Stocklin R, Menin L. Anti-microbial peptides: from invertebrates tovertebrates. Immunological Reviews,2004,198:169~84
[11] Butler R A, Kelley M L, Olberdingb K E, et al. Aryl hydrocarbon receptor(AhR)-independent effects of2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) onsoftshell clam (Mya arenaria) reproductive tissue. Comparative Biochemistry andPhysiology Part C,2004,138:375~381
[12] Butler R A, Kelley M L, Powell W H, et al. An aryl hydrocarbon receptor (AHR)homologue from the soft-shell clam, Mya arenaria: evidence that invertebrateAHR homologues lack2,3,7,8-tetrachlorodibenzo-p-dioxin andβ-naphthoflavone binding. Gene,2001,278:223~234
[13] Canova S, Degan P, Peters L D, et al. Tissue dose, DNA adducts, oxidative DNAdamage and CYP1A-immunopositive proteins in mussels exposed to waterbornebenzo[a]pyrene. Mutation Research,1998,399:17~30
[14] Carr R S, Biedenbach J M, Hooten R I. Sediment quality assessment survey andtoxicity identification evaluation studies in Lavaca Bay, Texas, a marinesuperfund site. Environmental Toxicology,2001,16(1):20~30
[15] Carver L A, Bradfield C A. Ligand-dependent interaction of the aryl hydrocarbonreceptor with a novel immunophilin homolog in vivo. Journal of BiologicalChemistry,1997,272(17):11452~11456
[16] Charlet M, Chernysh S, Philippe H, et al. Isolation of several cysteine-richantimicrobial peptides from the blood of a mollusc, Mytilus edulis. Journal ofBiological Chemistry,1996,271:21808~21813
[17] Chaty S, Rodius F, Vasseur P. A comparative study of the expression of CYP1Aand CYP4genes in aquatic invertebrate (freshwater mussel, Unio tumidus) andvertebrate (rainbow trout, Oncorhynchus mykiss). Aquatic Toxicology,2004,69:81~93
[18] Cheung C C C, Siu W H L, Richardson S B, et al. Antioxidant responses tobenzo[a]pyrene and Aroclor1254exposure in the green-lipped mussel, Pernaviridis. Environmental Pollution,2004,128:393~403
[19] Cheung C C C, Zheng G J, Li A M Y. Relationships between tissueconcentrations of polycyclicaromatic hydrocarbons and antioxidative responsesof marine mussels, Perna viridis. Aquatic Toxicology,2001,52:189～203
[20] Diatchenko L, Lau Y F C, Campbell A P. Suppression subtractive hybridization:A method for generating differentially regμlated or tissue-specific cDNA probesand libraries. Proceedings of the National Academy of Sciences of the UnitedStates of America,1996,93:6025~6030
[21] Diatchenko L, Lukyanov S, Lau Y F C, et al. Suppression subtractivehybridization: a versatile method for identifying differentially expressed genes.Methods in Enzymology,1999,303:349~380
[22] Ding Y, Miao J L, Li G Y, et al. Efect of Cd on GSH and GSH-related enzymesof Chlamydomonas sp. ICE-L existing in Antarctic ice. Journal of EnvironmentalSciences,2005,17(4):667~67l
[23] Dolwick K M, Swanson H I, Bradfeld C A. In vitro analysis of Ah receptordomains involved in ligand-activated DNA recognition. Proceedings of theNational Academy of Sciences of the United States of America,1993,90:8566~8570
[24] Doyen P, Vasseur P, Rodius F. cDNA cloning and expression pattern of pi-classglutathione S-transferase in the freshwater bivalves Unio tumidus and Corbiculafluminea. Comparative Biochemistry and Physiology Part C,2005,140:300~308
[25] Drutel G, Kathmann M, Heron Anne, et al. Cloning and selective expression inbrain and kidney of ARNT2homologous to the Ah Receptor NuclearTranslocator (ARNT). Biochemical and Biophysical Research Communications,1996,225:333~339
[26] Duncan D M, Burgess E A, Duncan I. Control of distal antennal identity andtarsal development in Drosophila by spineless-aristapedia, a homolog of themammalian dioxin receptor. Genes&Development,1998,12:1290~1303
[27] Elnemr A, Abd-allan A M. Contamination of polycyclic aromatic hydrocarbons(PAHs) in microlayer and subsurface waters along Alexandria Coast, Egypt.Chemosphere,2003,52(10):711~1716
[28] Emmons R B, Duncan D, Estes P A, et al. The spineless-aristapedia and tangobHLH-PAS proteins interact to control antennal and tarsal development inDrosophila. Development,1999,126:3937~3945
[29] Fukunaga B N, Hankinson O. Identifcation of a novel domain in the arylhydrocarbon receptor required for DNA binding. Journal of Biological Chemistry,1996,271:3743~3749
[30] Fukunaga B N, Probst M R, Reiszporszasz S, et al. Identifcation of functionaldomains of the aryl hydrocarbon receptor. Journal of Biological Chemistry,1995,270:29270~29278
[31] Gao Q, Song L S, Ni D J, et al. cDNA cloning and mRNA expression of heatshock protein90gene in the haemocytes of Zhikong scallop Chlamys farreri.Comparative Biochemistry and Physiology Part B,2007,147:704~715
[32] Genomic Environmental Project. http://www. niehs. nih. gov/envgenom/home.htm.
[33] Gestal C, Costa M, Figueras A, et al. Analysis of differentially expressed genes inresponse to bacterial stimulationin hemocytes of the carpet-shell clam Ruditapesdecussatus: Identification of new antimicrobial peptides. Gene,2007,406:134~143
[34] Goksoyr A. Biomarker responses in flounder (Platichthys flesus) and their use inpollution monitoring. Marine Pollution Bulletin,1999,33(1-6):36~45
[35] Gonzalez M, Gueguen Y, Desserre G, et al. Molecular characterization of twoisoforms of defensin from hemocytes of the oyster Crassostrea gigas.Developmental&Comparative Immunology,2007,31(4):332~339
[36] Grote M, Schuurmann G, Altenburger R. Modeling Photoinduced Algal Toxicityof Polycyclic Aromatic Hydrocarbons. Environmental Science&Technology,2005,39:4141~4149
[37] Hankinson O. The aryl hydrocarbon receptor comples. Annual Review ofPharmacology and Toxicology,1995,35:307~340
[38] Hanssona M C, Wittzell H K, Perssona K, et al. Characterization of two distinctaryl hydrocarbon receptor (AhR2) genes in Atlantic salmon (Salmo salar) andevidence for multiple AhR2gene lineages in salmonid fish. Gene,2003,303:197~206
[39] Harper P A, Riddick D S, Okey A B. Regulating the regulator: Factors thatcontrol levels and activity of the aryl hydrocarbon receptor, BiochemicalPharmacology,2006,72:267~279
[40] He N H, Qin Q W, Xu X. Differential profile of genes expressed in hemocytes ofWhite Spot Syndrome Virus-resistant shrimp (Penaeus japonicus) by combiningsuppression subtractive hybridization and differential hybridization. AntiviralResearch,2005,66:39~45
[41] Hoarau P, Damiens G, Roméo M, et al. Cloning and expression of a GST-pi genein Mytilus galloprovincialis. Attempt to use the GST-pi transcript as a biomarkerof pollution. Comparative Biochemistry and Physiology Part C,2006,143:196~203
[42] Holley S J, Yamamoto K R. A role for HSP90in retinoid receptor signaltransduction. Molecular and Cellular Biology,1995,6:1833~1842
[43] Holtha T F, Nourizadeh-Lillabadic R, Blaesbjergd M, et al. Differential geneexpression and biomarkers in zebrafish (Danio rerio) following exposure toproduced water components. Aquatic Toxicology,2008,90:277~291
[44] Hook S E, Cobb M E, Oris J T, et al. Gene sequences for Cytochromes p4501A1and1A2: The need for biomarker development in sea otters (Enhydra lutris).Comparative Biochemistry and Physiology Part B,2008,151:336~348
[45] Hubank M, Schatz D. Identifiying difference in mRNA expression byrepresentational difference analysis of cDNA. Nucleic Acids Research,1994,22:5640~5648
[46] Hubert F, No l T, Roch P. A member of the arthropod defensin family from theedible Mediterranean mussels (Mytilus galloprovincialis). European Journal ofBiochemistry,1996,240:302~306
[47] Hulka B S, Garrett P. Influence of biomarkers on the design, methods, andinterpretation of epidemiologic studies. Journal of Toxicology and EnvironmentalHealth,1993,40(2-3):389
[48] Iijima R, Kisugi J, Yamazaki M. A novel antimicrobial peptide from the sea hareDolabella auricularia. Developmental and Comparative Immunology,2003,27:305~311
[49] Jain S, Dolwick K M, Schmidt J V, et al. Potent transactivation domains of theAh receptor and Ah receptor nuclear translocator map to their carboxyl termini.Journal of Biological Chemistry,1994,269:31518~31524
[50] Ji W, Wright M B, Cai L, et al. Efficacy of SSH PCE in isolating differentiallyexpressed genes. BMC Genomics,2002,3:12~16
[51] Kann S, Huang M Y, Estes C, et al. Arsenite-induced aryl hydrocarbon receptornuclear translocation results in additive induction of phase I genes and synergisticinduction of phase II genes. Molecular Pharmacology,2005,68:336~346
[52] Lake J, Gravel C, Koko G K, et al. Combining suppressive subtractivehybridization and cDNA microarrays to identify dietary phosphorus-responsivegenes of the rainbow trout (Oncorhynchus mykiss) kidney. ComparativeBiochemistry and Physiology Part D,2010,5(1):24~35
[53] Lang Q, Zhang Q, Jaffe R. Organic aerosols in the Miami area, USA: temporalvariability of atmospheric particlesand wet/dry deposition. Chemosphere,2002,47(4):427~441
[54] Lange B M, Bachi A, Wilm M, et al. HSP90is a core centrosomal componentand is required at different stages of the centrosome cycle in Drosophila andvertebrates. Embo Journal,2000,19:1252~1262
[55] Lee J S, Kim E Y, Iwata H, et al. Molecular characterization and tissuedistribution of aryl hydrocarbon receptor nuclear translocator isoforms, ARNT1and ARNT2, and identification of novel splice variants in common cormorant(Phalacrocorax carbo). Comparative Biochemistry and Physiology Part C,2007,145:379~393
[56] Leelatanawit R, Klinbunga S, Aoki T. Suppression subtractive hybridization(SSH) for isolation and characterization of genes related to testiculardevelopment in the giant tiger shrimp Penaeus monodon. BMB Reports,2008,796~802
[57] Li C, Ni D, Song L, et al. Molecular cloning and characterization of a catalasegene from Zhikong scallop Chlamys farreri. Fish&shellfish immunology,2008,24(1):26~34
[58] Liang P, Pardee A. Differential display of eukaryotic messenger RNA by meansof the polymerase chain reaction. Science,1992,257:967~971
[59] Lisitsyn N, Wigler M. Cloning the differences between two complex genomes.Science,1993,259:946~951
[60] Livak K J, Schmittgen T D. Analysis of relative gene expression data usingreal-time quantitative PCR and the2(-Delta Delta C(T)) Method. Methods,2001,25:402~408
[61] Luedeking A, Koehler A. Regulation of expression of multixenobiotic resistance(MXR) genes by environmental factors in the blue mussel Mytilus edulis. AquaticToxicology,2004,69:1~10
[62] Magi E, Bianco R, Ianni C, et al. Distribution of polycyclic aromatichydrocarbons in the sediments of the Adriatic Sea. Environmental Pollution,2002,119(1):91~98
[63] Maliszewska K B. Dissipation of polycyclic aromatic hydrocarbons in freshlycontaminated soils-the effect of soil physicochemical p roperties and aging. Water,Air and Soil Pollution,2005,168:113~128
[64] Marchler-Bauer A, Bryant S H. CD-Search: protein domain annotations on thefly. Nucleic Acids Research,2004,32(W):327~331
[65] Merson R R, Franks D G, Karchner S I, et al. Development and characterizationof polyclonal antibodies against the aryl hydrocarbon receptor protein family(AHR1, AHR2, and AHR repressor) of Atlantic killifish Fundulus heteroclitus.Comparative Biochemistry and Physiology Part C,2006,142:85~94
[66] Nahrganga J, Camusa L, Gonzalezc P, et al. PAH biomarker responses in polarcod (Boreogadus saida) exposed to Benzo(a)pyrene. Aquatic Toxicology,2009,94:309~319
[67] Nebert D W, Roe A L, Dieter M Z, et al. Role of the aromatic hydrocarbonreceptor and [Ah]gene battery in the oxidative stress response, cell cycle control,and apotosis. Biochemical Pharmacology,2000,59(1):65~85
[68] Nebert D W. The Ah locus: genetic differences in toxicity, cancer, mutation, andbirth defects. Critical Reviews in Toxicology,1989,20(3):153~174
[69] Necela B, Pollenz R S. Functional analysis of activation and repression domainsof the rainbow trout Aryl hydrocarbon receptor nuclear translocator (rtARNT)protein isoforms. Biochemical Pharmacology,1999,57:1177~1190
[70] Ni D J, Song L S, Gao Q, et al. The cDNA cloning and mRNA expression ofcytoplasmic Cu, Zn superoxide dismutase (SOD) gene in scallop Chlamys farreri.Fish&Shellfish Immunology,2007,23(5):1032~1042
[71] Notar M, Leskovsek H, Faganeli J. Composition, distribution and sources ofpolycyclic aromatic hydrocarbons in sediments of the Gulf of Trieste, NorthernAdriatic Sea. Marine Pollution Bulletin,2001,1:36~44
[72] Ohi H, Fujita Y, Miyao M et al. Molecular cloning and expression analysis of thearyl hydrocarbon receptor of Xenopus laevis. Biochemical and BiophysicalResearch Communications,2003,307:595~599
[73] Oost R, Beyer J, Vermeulen N P E. Fish bioaccumulation and biomarkers inenvironmental risk assessment: a review. Environmental Toxicology andPharmacology,2003,13:57~149
[74] Paine M D, Chapman P M, Allard P J. Limited bioavailability of sediment PAHnear an aluminium smelter: contamination does not equal effects. EnvironmentalToxicology and Chemistry,1996,15:2003~2018
[75] Pan L Q, Liu N, Xu C Q, et al. Identification and expression of a novelcytochrome P450gene, belong to CYP4, in the clam Ruditapes philippinarumexposed to Benzo(a)pyrene. Environmental Toxicology and Pharmacology,2011,32:390~398
[76] Pandey P, Saleh A, Nakazawa A, et al. Negative regulation of cytochromeC-mediated oligomerization of Apaf-1and activation of procaspase~9by heatshock protein90. Embo Journal,2000,19:4310~4322
[77] Pandit G G, Sahu S K, Puranik V D. Exchange of polycyclic aromatichydrocarbons across the air-water interface at the creek adjoining MumbaiHarbour, India. Environment International,2006,32(2):259~264
[78] Pascual P, Pedrajas J R,Toribi F, et al. Effect of food deprivation on oxidativestress biomarke in fish (sparus aurata). Chemico-Biological Interactions,2003,145:191~199
[79] Paul C, Hartmann J C. The distribution and sources of polycyclic aromatichydrocarbons in Narragansett Bay surface sediments. Marine Pollution Bulletin,2004,48(3/4):351~358
[80] Powell-Coffman J A, Bradfield C A, Wood A W. Caenorhabditis elegansorthologs of the aryl hydrocarbon receptor and its heterodimerization partner thearyl hydrocarbon receptor nuclear translocator. Biochemistry,1998,95:2844~2849
[81] Ross J R, Oros D R. Polycyclic aromatic hydrocarbons in the San FranciscoEstuary water column: sources, spatial distributions, and temporal trends(1993—2001). Chemosphere,2004,57(8):909~920
[82] Saha S, Sparks A B, Rago C, et al. Using the transcriptomet to annotate thegeome. Nature Biotechnology,2002,20:508~512
[83] Sayeed I, Parvez S, Pandey S, et al. Oxidative stress biomarkers of exposure todeltamethrin in freshwaterfish, Channa punctatus Bloch. Ecotoxicology andEnvironmental Safety,2003,56:295~301.
[84] Schena M, Shalon D, Davis R, et al. Quantitative monitoring of gene expressionpatterns with a completementary DNA microarray. Science,1995,270:467~470
[85] Shi Y H, Liu J, Xia J H, et al. Screen for stage-specific expression genes betweentail bud stage and heartbeat beginning stage in embryogenesis of gynogeneticsilver crucian carp. Cell Research,2002,12(2):133~142
[86] Sicre M A, Marty J C, Saliot A. Aliphatic and aromatic hydrocarbons in theMediterranean aerosol. International Journal of Environmental AnalyticalChemistry,1987,29(1/2):73~94
[87] Snyder M J. Cytochrome P450enzymes belonging to the CYP4family frommarine invertebrates. Biochemical and Biophysical Research Communications,1998,249:187~190
[88] Stefan B, Anneli W. Photochemical degradation of PAHs in freshwaters and theirimpact on bacterial growth influence of water chemistry. Hydrobiologia,2002,469:23~32
[89] Swanson H I, Bradfield C A. The AH-receptor: genetics, structure and function.Pharmacogenetics,1993,3(5):213~230
[90] Swanson H I, Yang J H. Mapping the protein/DNA contact sites of the Ahreceptor and Ah receptor nuclear translocator. Journal of Biological Chemistry,1996,271:31657~31665
[91] Tanguy A, Guo X M, Ford S E. Discovery of genes expressed in response toPerkinsus marinus challenge in Eastern (Crassostrea virginica) and Pacific (C.gigas) oysters. Gene,2004,338:121~131
[92] Telli-Karakoc F, Tolun L, Henkelmann B. Polycyclic aromatic hydrocarbons(PAHs) and polychlorinated biphenyls (PCBs) distributions in the Bay ofMarmara Sea: lzmit Bay. Environmental Pollution,2002,119(3):383~397
[93] Tolosa I, Demora S, Sheikholeslami M R, et al. Aliphatic and aromatichydrocarbons in coastal Caspian Sea sediments. Marine Pollution Bulletin,2004,48(1-2):44~60
[94] Toyooka T, Ibuli Y. New Method for Testing Phototoxicity of PolycyclicAromatic Hydrocarbons. Environmental Science&Technology,2006,40:3603~3608
[95] Tsai W T, Mi H H, Chang Y M, et al. Polycyclic aromatic hydrocarbons (PAHs)in biocrudes from induction-heating pyrolysis of biomass wastes. BioresourceTechnology,2007,98:1133~1137
[96] Vance E D, Brookes P C, Jenkinson D S. An extraction method for measuringsoil microbial biomass C. Soil Biology&Biochemistry,1987,19:703~707
[97] Vicguri J, Verde J, Irabien A. Environmental assessment of polycyclic aromatichydrocarbons (PAHs) in surface sediments of the Santander Bay, Northern Spain.Chemosphere,2002,48(2):157~165
[98] Wang B, Li F H, Dong B, et al. Discovery of the genes in response to white spotsyndrome virus (WSSV) infection in Fenneropenaeus chinensis through cDNAmicroarray. Marine Biotechnology,2006,8(5):491~500
[99] Wang K J, Ren H L, Xu D D, et al. Identification of the up-regulated expressiongenes in hemocytes ofvariously colored abalone (Haliotis diversicolor Reeve,1846) challenged with bacteria. Developmental and Comparative Immunology,2008,32:1326~1347
[100] Wang L, Pan LQ, Liu N, et al. Biomarkers and bioaccumulation of clamRuditapes philippinarum in response to combined cadmium and benzo[α]pyreneexposure. Food and Chemical Toxicology,2011,49:3407~3417
[101] Wang Z L, Wu Z H, Jian J C, et al. Cloning and expression of heat shockprotein70gene in the haemocytes of pearl oyster (Pinctada fucata, Gould1850)responding to bacterial challenge. Fish Shellfish Immunology,2009,26:639~645
[102] Whitelaw M, Gottlicher M, Gustafsson J A, et al. Defnition of a novel ligandbinding domain of a nuclear bHLH receptor: co-localization of ligand and HSP90binding activities within the regulable inactivation domain of the dioxinreceptor. Embo Journal,1993,12:4169~4179
[103] WHO. IPCS Environmental Health Criteria, Vol155: Biomarkers and riskassessment: concepts and principles [Z]. Geneva: World Health Organization,1993
[104] Wiech H, Buehner J, Zimmermann R, et al. HSP90chaperones protein foldingin vitro, Nature,1992,358:169~170
[105] Willett K L, Wilson C, Thomsen J, et al. Evidence for and against the presenceof polynuclear aromatic hydrocarbon and2,3,7,8-tetrachloro-p-dioxin bindingproteins in the marine mussels, Bathymodiolus and Modiolus moliolus. AquaticToxicology,1999,48:51~64
[106] Witt G, Leipe T, Emeis K C. Using fluffy layer material to study the fate ofparticle-bound organic pollutants in the sourthern Baltic Sea. EnvironmentalScience&Technology,2001,35(8):1567~1573
[107] Witt G. Occurrence and transport of polycyclic aromatic hydrocarbons in thewater bodies of Baltic Sea. Marine Chemistry,2002,79(2):49~66
[108] Xu C Q, Pan L Q, Liu N, et al. Cloning, characterization and tissue distributionof a pi-class glutathione S-transferase from clam (Venerupis philippinarum):Response to Benzo[α]pyrene exposure. Comparative Biochemistry andPhysiology Part C,2010,152:160~166
[109] Zhang H, Song L S, Li C H, et al. Molecular cloning and characterization of athioester-containing protein from Zhikong scallop Chlamys farreri. MolecularImmunology,2007,44:3492~3500
[110] Zhang J F, Liu H, Sun Y Y, et al. Responses of the antioxidant defenses of theGoldfish Carassius auratus, exposed to2,4-dichlorophenol. EnvironmentalToxicology and Pharmacology,2005,19:185~190
[111] Zhang L, Li F G, Liu C L, et al. Construction and analysis of cotton (Gossypiumarboreum L.) drought-related cDNA library. BMC Research Notes,2009,2:120
[112] Zhang Z, Wang Y, Jiang Y, et al. Ribosomal protein124is differentiallyexpressed in ovary and testis of themarine shrimpmar-supenaeus japonicus.Biochem and Molecular Biology,2007,147(3):466~474
[113] Zhou J L, Maskaoui K. Distribution of polycyclic aromatic hydrocarbons inwater and surface sediments from Daya Bay, China. Environmental Pollution,2003,121(2):269~281
[114]安社娟,2005.多环芳烃致癌的分子毒理学研究进展.国外医学卫生学分册,32(1):10~13
[115]百度百科:《生物监测》,2012年4月24日,http://baike.baidu.com/view/469702.htm.
[116]百度百科:《抑制性消减杂交技术》,2010年12月7日,http://baike.baidu.com/view/4494504.htm.
[117]包艳波,徐日福,王志贤,等.鸡卵巢组织SSH cDNA文库构建及差异表达基因筛选与初步鉴定.吉林农业大学学报,2011.(http://www.cnki.net/kcms/detail/22.1100.s.20110516.1625.001.html)
[118]曾勇一,陆承平. cDNA微阵列结合抑制性差减杂交研究对虾白斑综合征病毒部分表达基因.病毒学报,2004,20(3):255~260
[119]陈亮,郭红岩,沈红,等.低浓度铅暴露对鲫鱼肝脏抗氧化系统的影响.环境化学,2002,21(5):485~489
[120]陈荣,刘辉,李东晓,等.水生动物谷胱甘肽硫转移酶研究进展.厦门大学学报自然科学版，2006,A02:176~184
[121]陈荣,郑微云,余群,等.石油污染对僧帽牡蛎(Ostrea cucullata)抗氧化酶的影响.环境科学学报,2002,22(3):385~388
[122]陈胜,董美阶,朱海燕,等.接触苯工人血浆热应激蛋白70抗体滴度分析.中华预防医学杂志,2000,34(1):8~10
[123]陈韬,苏建明,文祝友,章怀云,2003.扇贝性腺和肠组织表达序列标签的初步识别.农业生物技术学报.11(4):407~411
[124]单晓枫,孙佳敏,张洪波,等.抑制性消减杂交技术及其在水生动物基因克隆中的应用.黑龙江畜牧兽医,2010,12:30~32
[125]董瑞斌,许东风,刘雷,等.多环芳烃在环境中的行为.环境与开发,1999,14(2):10~11
[126]范志杰,宋春印.海洋环境污染生物监测及其方案的设计研究．交通环保，1994,1:3~8.
[127]方孝东,黄薇,林栖凤,等.通过cDNA RDA法分离和识别盐藻(Dunaliella salina)盐胁迫相关基因.中国生物化学与分子生物学报,2004,20(1):67~72
[128]韩菲,2007.多环芳烃来源与分布及迁移规律研究概述.气象与环境学报,23(4):57~61
[129]黄春,李润琴,张万江,2009.应用抑制性消减杂交技术筛选结核分枝杆菌差异基因.中华微生物学和免疫学杂志.6:507~512
[130]黄耀芬,李丽萍,胡显安,李再培.水质有机污染的指示生物——寡毛类.黑龙江环境通报,2003,2:71~72
[131]霍传林.近海环境中有机污染物对底栖鱼体内EROD活性的诱导作用研究:
[硕士学位论文].大连:大连理工大学.2003
[132]基因时代:《基因表达序列分析（SAGE）》,2009年9月14日,http://ggene.cn/html/protocol/bioinformatics/2009/0914/3126.html.
[133]吉成龙,孙国华,杨建敏,等.刺参(Apostichopus japonicus)高温胁迫消减cDNA文库的构建与分析.海洋与湖沼,2011,1:62~68
[134]江桂斌.持久性有毒污染物的环境化学行为与毒理效应.毒理学杂志,2005,19(3):179~180
[135]姜淑梅,吕晓梅,胡赞民.差减抑制杂交技术在植物研究中的应用.植物学通报,2005,22(增刊):91~98
[136]金赞芳,陈英旭.环境的PAHs污染及其生物修复技术研究进展.农业环境保护,2001,20(2):123~125
[137]孔繁祥.环境生物学.北京:高等教育出版社,2000.51-56
[138]匡少平,孙东亚.多环芳烃的毒理学特征与生物标记物研究.世界科技研究与发展,2007,29(2):41~47
[139]郎印海,贾永冈,刘宗峰,等.黄河口水中多环芳烃(PAHs)的季节分布特征及来源分析.中国海洋大学学报,2008,38(4):640~646
[140]李立芹,鲁黎明,2006.抑制性消减杂交技术（SSH）及其在烟草生物学研究中的应用.中国烟草科学.32(3):89~94
[141]李文雍,郁卫东,刘桂生,等.用DDRT—PCR技术克隆小鼠早期胚胎发育相关基因.中国生物化学与分子生物学报,2002, l8(6):728~732
[142]李小庆,景志忠.抑制性消减杂交技术的应用.生物技术通报,2009,5:46~50
[143]林道辉,朱利中,王静.小冶炼地区PAHs污染及其风险评价.生态学报,2005,25(2):262~267
[144]凌晰,刘文新,陈江麟,等.黄海近岸表层沉积物中PAHs的分布特征与潜在风险.环境科学学报,2008,28(7):1395~1399
[145]刘海芳,王凡.水环境污染程度的分子生物标志物的研究进展.安徽农业科学,2007,35(18):5502,5503~5506
[146]刘辉,叶湘漓.文昌鱼抗金葡菌感染差减文库的构建及免疫相关基因分析.中国生物化学与分子生物学报,2009,25(8):727~733
[147]刘晶,张文兵,麦康森,等.皱纹盘鲍外套膜耐维生素E缺乏消减cDNA文库的构建.中国水产科学,2007,14(3):383~389
[148]刘静.苯并a芘对栉孔扇贝血细胞免疫毒性机制的研究:[博士学位论文].青岛:中国海洋大学.2009
[149]刘玲.环芳烃的代谢和肺癌易感性的研究.国外医学卫生学分册,1986,3:136~140
[150]刘维全,高士争,王吉贵.分子生物学实验基本操作技术.北京:化学工业出版社,2010-04
[151]刘岩,张祖麟,洪华生.厦门西港表层海水PAHs含量分布特征和来源进行分析.海洋通报,1999,8(4):38~43
[152]刘移民.芳香烃类化合物受体研究进展.中国职业医学,2001,28(6):46~48
[153]马明辉,海志皆,冯志权,等.辽东湾双台子河海区动物体内污染物含量及时空分布.海洋环境科学,1999,18(1):61~76
[154]马伟业,千新来,赵清正,等.应用抑制性消减杂交技术分离E1A药敏相关基因.癌症,2004,23(2):146~149
[155]马长艳,周开亚,郭豫杰,等.中华绒螯蟹卵巢差减cDNA文库的构建.动物学研究,2003,24(1):53~56
[156]麦碧娴,林峥,张干,等.珠江三角洲沉积物中毒害有机物的污染现状及评价.环境科学研究,2001,14(1):19~23
[157]麦碧娴，林峥，张干,等.珠江三角洲河流和珠江口表层沉积物中有机污染物研究一多环芳烃和有机氯农药的分布及特征.环境科学学报,2000,2(2):192~197
[158]苗晶晶.苯并[a]芘对栉孔扇(Chlamys farreri)贝分子毒理学机制的研究:[博士学位论文].青岛:中国海洋大学,2010
[159]潘鲁青,刘娜,王静.栉孔扇贝在B[a]P胁迫下生物标志物筛选的研究.水生生物学报,2012,36(2):1~8
[160]庞朋沙,过倩萍,伍会健.细胞内AhR信号转导通路的机制研究.现代生物医学进展,2010,10(13):2567~2570
[161]戚元成,马雷,王菲菲,等.打顶烟草根尖组织抑制性消减cDNA文库的构建及其序列分析.中国农业科学,2011,44(7)
[162]丘耀文,周俊良, Maskaoui K.,等.大亚湾海域水体和沉积物中多环芳烃分布及其生态危害评价,2004,23(4):72~80
[163]曲跃军,杜人杰,王雷,等.抑制性消减杂交技术研究Cd2+胁迫下星星草基因表达.东北林业大学学报,2010,38(2):9~11
[164]饶志明,董海涛,庄杰云,等.水稻稻瘟病菌胁迫应答cDNA片段的表达及定位.中国水稻科学,2003,17(2):105~108
[165]任加云,潘鲁青,苗晶晶.苯并(a)芘和苯并(k)荧蒽混合物对栉孔扇贝毒理学指标的影响.环境科学学报,2006,26(7):1180~1186
[166]任天志.持续农业中的土壤生物指标研究.中国农业科学,2000,33(1):68~75
[167]任秀华,斯陆勤,曹磊,等.聚氧乙烯醚类表面活性剂对大鼠体内细胞色素P4503A活性的影响.药学学报,2008,43(5):528~534
[168]生物秀:《传统mRNA差异显示技术与第二代差异显示系统》,2005年11月30日, http://www.bbioo.com/experiment/12-926-1.html.
[169]苏建明.草鱼肠道组织消减cDNA文库的构建及免疫相关基因的克隆与表达分析:[博士学位论文].长沙:湖南农业大学,2007
[170]唐中林,万平,李小平,等.猪胚胎骨骼肌LongSAGE文库构建.农业生物技术学报,2007,15(1):171~172
[171]滕彦国,倪师军,张成江,等.攀枝花地区不同工业区表层土壤中重金属分布的特征.物探化探计算技术,2002,24(3):254~262
[172]田蕴,郑天凌,王新红.厦门西港表层沉积物中多环芳烃(PAHs)的含量、分布及来源.海洋与湖沼,2004,35(1):16~20
[173]田蕴,郑天凌,王新红.厦门西港表层海水中PAHs的含量、组成及来源.环境科学学报,2004,24(1):50~55
[174]王春光,李桂兰,尹松年.苯染毒小鼠体内DNA加合物及其持续时间的研究.中华预防医学杂志,1993,11:344~345
[175]王静,潘鲁青,苗晶晶,2007.多环芳烃化合物对栉孔扇贝组织芳烃羟化酶活力的影响.海洋科学.31(12):19~23
[176]王伟,2009. PLC法测定水中16种多环芳烃.环境科学与管理,34(7):108~111
[177]王晓蓉,罗义,施华宏,等.分子生物标志物在污染环境早期诊断和生态风险评价中的应用.环境化学,2006,25(3):320~325
[178]王秀丽,徐建民,姚槐应,等.重金属铜、锌、镉、铅复合污染对土壤环境微生物群落的影响.环境科学学报,2003,23(1):22~27
[179]王学彤,贾英,孙阳昭,等.典型污染区河流表层沉积物中PAHs的分布来源及生态风险.环境科学,2010,31(1):153~158
[180]王在照,焦传珍,张晓军,等.中国对虾蜕皮抑制激素全长cDNA的克隆及序列分析.遗传学报,2003,30(2):12~134
[181]王卓,张万起.芳香烃受体核转位蛋白的结构及相关功能.生命科学,2007,19(1):73~77
[182]徐凤山,张均龙.中国海典型生境双壳类软体动物多样性特点.生物多样性,2011,19(6):716~722
[183]许光辉,郑洪元.土壤微生物分析方法手册[M].上海:上海科技出版社,1986.226~228
[184]闫博,王兰,李涌泉,等.镉对长江华溪蟹肝胰腺抗氧化酶活力的影响.动物学报,2007,53(6):1121~1128
[185]阳晶.沙田柚花柱自交不亲和相关基因的克隆与鉴定:[硕士学位论文].桂林:广西师范大学,2008
[186]杨佰娟,郑立,陈军辉,等.南黄海中部表层沉积物中多环芳烃含量分布及来源分析,2009,29(3):663~667
[187]杨涛,陈海刚,蔡文贵,等.菲和苯并荧蒽曝露对翡翠贻贝外套膜的氧化胁迫及损伤.南方水产科学,2011,7(4):24~29
[188]杨毅,刘敏,侯立军,等.海岸带水环境中多环芳烃的归宿研究.海洋环境科学,2003,22(1):69~72
[189]杨志新,刘树庆.重金属Cd、Zn、Pb复合污染对土壤酶活性的影响.环境科学学报,2001,21(1):60~63
[190]叶细标,倪为民,傅华.分子生物标志物及其应用.中国工业医学杂志,2002,15(1):32~34
[191]于莹,张昌武,赵楠,等.胶州湾浮游纤毛虫丰度和生物量的周年变化.海洋与湖沼,2011,42(5):690~701
[192]袁东星,杨东宁,陈猛,等.厦门西港及闽江口表层沉积物中多环芳烃和有机氯污染物的含量及分布.环境科学学报，2001，21(1)：107~112
[193]岳桂东.玉米干旱胁迫相关基因的克隆与分析:[博士学位论文].济南:山东大学,2008
[194]岳敏,谷学新,邹洪.多环芳烃的危害与防治.首都师范大学学报,2003,24:40~44
[195]张娇.黄河及河口烃类有机物的分布特征及源解析:[博士学位论文].青岛:中国海洋大学,2008
[196]张强,张志文,龚侃,等.应用抑制性消减杂交技术克隆鉴定肾癌特异表达基因.科学通报,2000,45(16):1758~1762
[197]张先勇,刁晓平,杨宝，等.苯并[a]芘对马氏珠母贝血淋巴细胞DNA损伤的研究.生态环境学报,2011,20(6-7):1107~1110
[198]张新庆,杨佰娟,黎先春,等.南黄海中部海水中多环芳烃的分布特征.岩矿测试,2009,28(4):325~328
[199]张义兵,石耀华,桂建芳.鱼类培养细胞抗病毒基因差减cDNA文库的构建.水生生物学报,2003,27(2):113~118
[200]章卓,雷红宇,苏建明,2010.草鱼肠道组织抑制性消减表达序列标签的初步分析.湖南农业大学学报(自然科学版).36(3):339~343
[201]周驰,李纯厚,张为民，等.翡翠贻贝CYP4基因克隆及其表达水平分析.热带海洋学报,2010,29(4):82~88
[202]朱婧,杜立新:《反转录差异显示技术(DDRT-PCR)及其方法的改进》，2008年7月19日, http://www.biomart.cn/article/9/26/27/5993.htm.
[203]庄志雄,张桥.分子生物标志物研究的现状与展望.中华劳动卫生职业病杂志,1997,15(3):131~133
[2] Altschul S F, Madden T L, Sch ffer A A, et al. Gapped BLAST and PSI-BLAST: anew generation of protein database search programs. Nucleic Acids Research,1997,25(17):3389~3402
[3] Atchley W, Fitch W M. A natural classification of the basic helix–loop–helix classof transcription factors. Evolution,1997,94:5172~5176
[4] Berg P, Pongratz I. Differential Usage of nuclear export sequences regulatesintracellular localization of the Dioxin (Aryl Hydrocarbon) receptor. Journal ofBiological Chemistry,2001,276(46):43231~43238
[5] Bollérot K, Angelier N, Coumailleau P. Molecular cloning and embryonicexpression of the Xenopus Arnt gene. Mechanisms of Development,2001,108:227~231
[6] Booij K, Hoedemaker J R, Bakker J F. Dissolved PCEs, PAHs, and HCB in porewaters and overlying waters of contaminated harbor sediments. EnvironmentalScience&Technology,2003,37(18):4213~4220
[7] Boulanger N, Lowenberger C, Volf P, et al. Characterization of a defensin from thesand fly Phlebotomus duboscqi induced by challenge with bacteria or theprotozoan parasite Leishmania major. Infection and Immunity,2004,72:7140~7146
[8] Brown D J, Beneden R J V, Clark G C. Identification of two binding proteins forhalogenated aromatic hydrocarbons in the hard-shell clam, Mercenariamercenaria. Archives of Biochemistry and Biophysics,1995,319(1):217~224
[9] Brown M, Davies I M, Moffat C F, et al. Application of SSH and a macroarrayto investigate altered gene expression in Mytilus edulis in response to exposure tobenzo[a]pyrene. Marine Environmental Research,2006,62: S128~S135
[10] Bulet P, Stocklin R, Menin L. Anti-microbial peptides: from invertebrates tovertebrates. Immunological Reviews,2004,198:169~84
[11] Butler R A, Kelley M L, Olberdingb K E, et al. Aryl hydrocarbon receptor(AhR)-independent effects of2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) onsoftshell clam (Mya arenaria) reproductive tissue. Comparative Biochemistry andPhysiology Part C,2004,138:375~381
[12] Butler R A, Kelley M L, Powell W H, et al. An aryl hydrocarbon receptor (AHR)homologue from the soft-shell clam, Mya arenaria: evidence that invertebrateAHR homologues lack2,3,7,8-tetrachlorodibenzo-p-dioxin andβ-naphthoflavone binding. Gene,2001,278:223~234
[13] Canova S, Degan P, Peters L D, et al. Tissue dose, DNA adducts, oxidative DNAdamage and CYP1A-immunopositive proteins in mussels exposed to waterbornebenzo[a]pyrene. Mutation Research,1998,399:17~30
[14] Carr R S, Biedenbach J M, Hooten R I. Sediment quality assessment survey andtoxicity identification evaluation studies in Lavaca Bay, Texas, a marinesuperfund site. Environmental Toxicology,2001,16(1):20~30
[15] Carver L A, Bradfield C A. Ligand-dependent interaction of the aryl hydrocarbonreceptor with a novel immunophilin homolog in vivo. Journal of BiologicalChemistry,1997,272(17):11452~11456
[16] Charlet M, Chernysh S, Philippe H, et al. Isolation of several cysteine-richantimicrobial peptides from the blood of a mollusc, Mytilus edulis. Journal ofBiological Chemistry,1996,271:21808~21813
[17] Chaty S, Rodius F, Vasseur P. A comparative study of the expression of CYP1Aand CYP4genes in aquatic invertebrate (freshwater mussel, Unio tumidus) andvertebrate (rainbow trout, Oncorhynchus mykiss). Aquatic Toxicology,2004,69:81~93
[18] Cheung C C C, Siu W H L, Richardson S B, et al. Antioxidant responses tobenzo[a]pyrene and Aroclor1254exposure in the green-lipped mussel, Pernaviridis. Environmental Pollution,2004,128:393~403
[19] Cheung C C C, Zheng G J, Li A M Y. Relationships between tissueconcentrations of polycyclicaromatic hydrocarbons and antioxidative responsesof marine mussels, Perna viridis. Aquatic Toxicology,2001,52:189～203
[20] Diatchenko L, Lau Y F C, Campbell A P. Suppression subtractive hybridization:A method for generating differentially regμlated or tissue-specific cDNA probesand libraries. Proceedings of the National Academy of Sciences of the UnitedStates of America,1996,93:6025~6030
[21] Diatchenko L, Lukyanov S, Lau Y F C, et al. Suppression subtractivehybridization: a versatile method for identifying differentially expressed genes.Methods in Enzymology,1999,303:349~380
[22] Ding Y, Miao J L, Li G Y, et al. Efect of Cd on GSH and GSH-related enzymesof Chlamydomonas sp. ICE-L existing in Antarctic ice. Journal of EnvironmentalSciences,2005,17(4):667~67l
[23] Dolwick K M, Swanson H I, Bradfeld C A. In vitro analysis of Ah receptordomains involved in ligand-activated DNA recognition. Proceedings of theNational Academy of Sciences of the United States of America,1993,90:8566~8570
[24] Doyen P, Vasseur P, Rodius F. cDNA cloning and expression pattern of pi-classglutathione S-transferase in the freshwater bivalves Unio tumidus and Corbiculafluminea. Comparative Biochemistry and Physiology Part C,2005,140:300~308
[25] Drutel G, Kathmann M, Heron Anne, et al. Cloning and selective expression inbrain and kidney of ARNT2homologous to the Ah Receptor NuclearTranslocator (ARNT). Biochemical and Biophysical Research Communications,1996,225:333~339
[26] Duncan D M, Burgess E A, Duncan I. Control of distal antennal identity andtarsal development in Drosophila by spineless-aristapedia, a homolog of themammalian dioxin receptor. Genes&Development,1998,12:1290~1303
[27] Elnemr A, Abd-allan A M. Contamination of polycyclic aromatic hydrocarbons(PAHs) in microlayer and subsurface waters along Alexandria Coast, Egypt.Chemosphere,2003,52(10):711~1716
[28] Emmons R B, Duncan D, Estes P A, et al. The spineless-aristapedia and tangobHLH-PAS proteins interact to control antennal and tarsal development inDrosophila. Development,1999,126:3937~3945
[29] Fukunaga B N, Hankinson O. Identifcation of a novel domain in the arylhydrocarbon receptor required for DNA binding. Journal of Biological Chemistry,1996,271:3743~3749
[30] Fukunaga B N, Probst M R, Reiszporszasz S, et al. Identifcation of functionaldomains of the aryl hydrocarbon receptor. Journal of Biological Chemistry,1995,270:29270~29278
[31] Gao Q, Song L S, Ni D J, et al. cDNA cloning and mRNA expression of heatshock protein90gene in the haemocytes of Zhikong scallop Chlamys farreri.Comparative Biochemistry and Physiology Part B,2007,147:704~715
[32] Genomic Environmental Project. http://www. niehs. nih. gov/envgenom/home.htm.
[33] Gestal C, Costa M, Figueras A, et al. Analysis of differentially expressed genes inresponse to bacterial stimulationin hemocytes of the carpet-shell clam Ruditapesdecussatus: Identification of new antimicrobial peptides. Gene,2007,406:134~143
[34] Goksoyr A. Biomarker responses in flounder (Platichthys flesus) and their use inpollution monitoring. Marine Pollution Bulletin,1999,33(1-6):36~45
[35] Gonzalez M, Gueguen Y, Desserre G, et al. Molecular characterization of twoisoforms of defensin from hemocytes of the oyster Crassostrea gigas.Developmental&Comparative Immunology,2007,31(4):332~339
[36] Grote M, Schuurmann G, Altenburger R. Modeling Photoinduced Algal Toxicityof Polycyclic Aromatic Hydrocarbons. Environmental Science&Technology,2005,39:4141~4149
[37] Hankinson O. The aryl hydrocarbon receptor comples. Annual Review ofPharmacology and Toxicology,1995,35:307~340
[38] Hanssona M C, Wittzell H K, Perssona K, et al. Characterization of two distinctaryl hydrocarbon receptor (AhR2) genes in Atlantic salmon (Salmo salar) andevidence for multiple AhR2gene lineages in salmonid fish. Gene,2003,303:197~206
[39] Harper P A, Riddick D S, Okey A B. Regulating the regulator: Factors thatcontrol levels and activity of the aryl hydrocarbon receptor, BiochemicalPharmacology,2006,72:267~279
[40] He N H, Qin Q W, Xu X. Differential profile of genes expressed in hemocytes ofWhite Spot Syndrome Virus-resistant shrimp (Penaeus japonicus) by combiningsuppression subtractive hybridization and differential hybridization. AntiviralResearch,2005,66:39~45
[41] Hoarau P, Damiens G, Roméo M, et al. Cloning and expression of a GST-pi genein Mytilus galloprovincialis. Attempt to use the GST-pi transcript as a biomarkerof pollution. Comparative Biochemistry and Physiology Part C,2006,143:196~203
[42] Holley S J, Yamamoto K R. A role for HSP90in retinoid receptor signaltransduction. Molecular and Cellular Biology,1995,6:1833~1842
[43] Holtha T F, Nourizadeh-Lillabadic R, Blaesbjergd M, et al. Differential geneexpression and biomarkers in zebrafish (Danio rerio) following exposure toproduced water components. Aquatic Toxicology,2008,90:277~291
[44] Hook S E, Cobb M E, Oris J T, et al. Gene sequences for Cytochromes p4501A1and1A2: The need for biomarker development in sea otters (Enhydra lutris).Comparative Biochemistry and Physiology Part B,2008,151:336~348
[45] Hubank M, Schatz D. Identifiying difference in mRNA expression byrepresentational difference analysis of cDNA. Nucleic Acids Research,1994,22:5640~5648
[46] Hubert F, No l T, Roch P. A member of the arthropod defensin family from theedible Mediterranean mussels (Mytilus galloprovincialis). European Journal ofBiochemistry,1996,240:302~306
[47] Hulka B S, Garrett P. Influence of biomarkers on the design, methods, andinterpretation of epidemiologic studies. Journal of Toxicology and EnvironmentalHealth,1993,40(2-3):389
[48] Iijima R, Kisugi J, Yamazaki M. A novel antimicrobial peptide from the sea hareDolabella auricularia. Developmental and Comparative Immunology,2003,27:305~311
[49] Jain S, Dolwick K M, Schmidt J V, et al. Potent transactivation domains of theAh receptor and Ah receptor nuclear translocator map to their carboxyl termini.Journal of Biological Chemistry,1994,269:31518~31524
[50] Ji W, Wright M B, Cai L, et al. Efficacy of SSH PCE in isolating differentiallyexpressed genes. BMC Genomics,2002,3:12~16
[51] Kann S, Huang M Y, Estes C, et al. Arsenite-induced aryl hydrocarbon receptornuclear translocation results in additive induction of phase I genes and synergisticinduction of phase II genes. Molecular Pharmacology,2005,68:336~346
[52] Lake J, Gravel C, Koko G K, et al. Combining suppressive subtractivehybridization and cDNA microarrays to identify dietary phosphorus-responsivegenes of the rainbow trout (Oncorhynchus mykiss) kidney. ComparativeBiochemistry and Physiology Part D,2010,5(1):24~35
[53] Lang Q, Zhang Q, Jaffe R. Organic aerosols in the Miami area, USA: temporalvariability of atmospheric particlesand wet/dry deposition. Chemosphere,2002,47(4):427~441
[54] Lange B M, Bachi A, Wilm M, et al. HSP90is a core centrosomal componentand is required at different stages of the centrosome cycle in Drosophila andvertebrates. Embo Journal,2000,19:1252~1262
[55] Lee J S, Kim E Y, Iwata H, et al. Molecular characterization and tissuedistribution of aryl hydrocarbon receptor nuclear translocator isoforms, ARNT1and ARNT2, and identification of novel splice variants in common cormorant(Phalacrocorax carbo). Comparative Biochemistry and Physiology Part C,2007,145:379~393
[56] Leelatanawit R, Klinbunga S, Aoki T. Suppression subtractive hybridization(SSH) for isolation and characterization of genes related to testiculardevelopment in the giant tiger shrimp Penaeus monodon. BMB Reports,2008,796~802
[57] Li C, Ni D, Song L, et al. Molecular cloning and characterization of a catalasegene from Zhikong scallop Chlamys farreri. Fish&shellfish immunology,2008,24(1):26~34
[58] Liang P, Pardee A. Differential display of eukaryotic messenger RNA by meansof the polymerase chain reaction. Science,1992,257:967~971
[59] Lisitsyn N, Wigler M. Cloning the differences between two complex genomes.Science,1993,259:946~951
[60] Livak K J, Schmittgen T D. Analysis of relative gene expression data usingreal-time quantitative PCR and the2(-Delta Delta C(T)) Method. Methods,2001,25:402~408
[61] Luedeking A, Koehler A. Regulation of expression of multixenobiotic resistance(MXR) genes by environmental factors in the blue mussel Mytilus edulis. AquaticToxicology,2004,69:1~10
[62] Magi E, Bianco R, Ianni C, et al. Distribution of polycyclic aromatichydrocarbons in the sediments of the Adriatic Sea. Environmental Pollution,2002,119(1):91~98
[63] Maliszewska K B. Dissipation of polycyclic aromatic hydrocarbons in freshlycontaminated soils-the effect of soil physicochemical p roperties and aging. Water,Air and Soil Pollution,2005,168:113~128
[64] Marchler-Bauer A, Bryant S H. CD-Search: protein domain annotations on thefly. Nucleic Acids Research,2004,32(W):327~331
[65] Merson R R, Franks D G, Karchner S I, et al. Development and characterizationof polyclonal antibodies against the aryl hydrocarbon receptor protein family(AHR1, AHR2, and AHR repressor) of Atlantic killifish Fundulus heteroclitus.Comparative Biochemistry and Physiology Part C,2006,142:85~94
[66] Nahrganga J, Camusa L, Gonzalezc P, et al. PAH biomarker responses in polarcod (Boreogadus saida) exposed to Benzo(a)pyrene. Aquatic Toxicology,2009,94:309~319
[67] Nebert D W, Roe A L, Dieter M Z, et al. Role of the aromatic hydrocarbonreceptor and [Ah]gene battery in the oxidative stress response, cell cycle control,and apotosis. Biochemical Pharmacology,2000,59(1):65~85
[68] Nebert D W. The Ah locus: genetic differences in toxicity, cancer, mutation, andbirth defects. Critical Reviews in Toxicology,1989,20(3):153~174
[69] Necela B, Pollenz R S. Functional analysis of activation and repression domainsof the rainbow trout Aryl hydrocarbon receptor nuclear translocator (rtARNT)protein isoforms. Biochemical Pharmacology,1999,57:1177~1190
[70] Ni D J, Song L S, Gao Q, et al. The cDNA cloning and mRNA expression ofcytoplasmic Cu, Zn superoxide dismutase (SOD) gene in scallop Chlamys farreri.Fish&Shellfish Immunology,2007,23(5):1032~1042
[71] Notar M, Leskovsek H, Faganeli J. Composition, distribution and sources ofpolycyclic aromatic hydrocarbons in sediments of the Gulf of Trieste, NorthernAdriatic Sea. Marine Pollution Bulletin,2001,1:36~44
[72] Ohi H, Fujita Y, Miyao M et al. Molecular cloning and expression analysis of thearyl hydrocarbon receptor of Xenopus laevis. Biochemical and BiophysicalResearch Communications,2003,307:595~599
[73] Oost R, Beyer J, Vermeulen N P E. Fish bioaccumulation and biomarkers inenvironmental risk assessment: a review. Environmental Toxicology andPharmacology,2003,13:57~149
[74] Paine M D, Chapman P M, Allard P J. Limited bioavailability of sediment PAHnear an aluminium smelter: contamination does not equal effects. EnvironmentalToxicology and Chemistry,1996,15:2003~2018
[75] Pan L Q, Liu N, Xu C Q, et al. Identification and expression of a novelcytochrome P450gene, belong to CYP4, in the clam Ruditapes philippinarumexposed to Benzo(a)pyrene. Environmental Toxicology and Pharmacology,2011,32:390~398
[76] Pandey P, Saleh A, Nakazawa A, et al. Negative regulation of cytochromeC-mediated oligomerization of Apaf-1and activation of procaspase~9by heatshock protein90. Embo Journal,2000,19:4310~4322
[77] Pandit G G, Sahu S K, Puranik V D. Exchange of polycyclic aromatichydrocarbons across the air-water interface at the creek adjoining MumbaiHarbour, India. Environment International,2006,32(2):259~264
[78] Pascual P, Pedrajas J R,Toribi F, et al. Effect of food deprivation on oxidativestress biomarke in fish (sparus aurata). Chemico-Biological Interactions,2003,145:191~199
[79] Paul C, Hartmann J C. The distribution and sources of polycyclic aromatichydrocarbons in Narragansett Bay surface sediments. Marine Pollution Bulletin,2004,48(3/4):351~358
[80] Powell-Coffman J A, Bradfield C A, Wood A W. Caenorhabditis elegansorthologs of the aryl hydrocarbon receptor and its heterodimerization partner thearyl hydrocarbon receptor nuclear translocator. Biochemistry,1998,95:2844~2849
[81] Ross J R, Oros D R. Polycyclic aromatic hydrocarbons in the San FranciscoEstuary water column: sources, spatial distributions, and temporal trends(1993—2001). Chemosphere,2004,57(8):909~920
[82] Saha S, Sparks A B, Rago C, et al. Using the transcriptomet to annotate thegeome. Nature Biotechnology,2002,20:508~512
[83] Sayeed I, Parvez S, Pandey S, et al. Oxidative stress biomarkers of exposure todeltamethrin in freshwaterfish, Channa punctatus Bloch. Ecotoxicology andEnvironmental Safety,2003,56:295~301.
[84] Schena M, Shalon D, Davis R, et al. Quantitative monitoring of gene expressionpatterns with a completementary DNA microarray. Science,1995,270:467~470
[85] Shi Y H, Liu J, Xia J H, et al. Screen for stage-specific expression genes betweentail bud stage and heartbeat beginning stage in embryogenesis of gynogeneticsilver crucian carp. Cell Research,2002,12(2):133~142
[86] Sicre M A, Marty J C, Saliot A. Aliphatic and aromatic hydrocarbons in theMediterranean aerosol. International Journal of Environmental AnalyticalChemistry,1987,29(1/2):73~94
[87] Snyder M J. Cytochrome P450enzymes belonging to the CYP4family frommarine invertebrates. Biochemical and Biophysical Research Communications,1998,249:187~190
[88] Stefan B, Anneli W. Photochemical degradation of PAHs in freshwaters and theirimpact on bacterial growth influence of water chemistry. Hydrobiologia,2002,469:23~32
[89] Swanson H I, Bradfield C A. The AH-receptor: genetics, structure and function.Pharmacogenetics,1993,3(5):213~230
[90] Swanson H I, Yang J H. Mapping the protein/DNA contact sites of the Ahreceptor and Ah receptor nuclear translocator. Journal of Biological Chemistry,1996,271:31657~31665
[91] Tanguy A, Guo X M, Ford S E. Discovery of genes expressed in response toPerkinsus marinus challenge in Eastern (Crassostrea virginica) and Pacific (C.gigas) oysters. Gene,2004,338:121~131
[92] Telli-Karakoc F, Tolun L, Henkelmann B. Polycyclic aromatic hydrocarbons(PAHs) and polychlorinated biphenyls (PCBs) distributions in the Bay ofMarmara Sea: lzmit Bay. Environmental Pollution,2002,119(3):383~397
[93] Tolosa I, Demora S, Sheikholeslami M R, et al. Aliphatic and aromatichydrocarbons in coastal Caspian Sea sediments. Marine Pollution Bulletin,2004,48(1-2):44~60
[94] Toyooka T, Ibuli Y. New Method for Testing Phototoxicity of PolycyclicAromatic Hydrocarbons. Environmental Science&Technology,2006,40:3603~3608
[95] Tsai W T, Mi H H, Chang Y M, et al. Polycyclic aromatic hydrocarbons (PAHs)in biocrudes from induction-heating pyrolysis of biomass wastes. BioresourceTechnology,2007,98:1133~1137
[96] Vance E D, Brookes P C, Jenkinson D S. An extraction method for measuringsoil microbial biomass C. Soil Biology&Biochemistry,1987,19:703~707
[97] Vicguri J, Verde J, Irabien A. Environmental assessment of polycyclic aromatichydrocarbons (PAHs) in surface sediments of the Santander Bay, Northern Spain.Chemosphere,2002,48(2):157~165
[98] Wang B, Li F H, Dong B, et al. Discovery of the genes in response to white spotsyndrome virus (WSSV) infection in Fenneropenaeus chinensis through cDNAmicroarray. Marine Biotechnology,2006,8(5):491~500
[99] Wang K J, Ren H L, Xu D D, et al. Identification of the up-regulated expressiongenes in hemocytes ofvariously colored abalone (Haliotis diversicolor Reeve,1846) challenged with bacteria. Developmental and Comparative Immunology,2008,32:1326~1347
[100] Wang L, Pan LQ, Liu N, et al. Biomarkers and bioaccumulation of clamRuditapes philippinarum in response to combined cadmium and benzo[α]pyreneexposure. Food and Chemical Toxicology,2011,49:3407~3417
[101] Wang Z L, Wu Z H, Jian J C, et al. Cloning and expression of heat shockprotein70gene in the haemocytes of pearl oyster (Pinctada fucata, Gould1850)responding to bacterial challenge. Fish Shellfish Immunology,2009,26:639~645
[102] Whitelaw M, Gottlicher M, Gustafsson J A, et al. Defnition of a novel ligandbinding domain of a nuclear bHLH receptor: co-localization of ligand and HSP90binding activities within the regulable inactivation domain of the dioxinreceptor. Embo Journal,1993,12:4169~4179
[103] WHO. IPCS Environmental Health Criteria, Vol155: Biomarkers and riskassessment: concepts and principles [Z]. Geneva: World Health Organization,1993
[104] Wiech H, Buehner J, Zimmermann R, et al. HSP90chaperones protein foldingin vitro, Nature,1992,358:169~170
[105] Willett K L, Wilson C, Thomsen J, et al. Evidence for and against the presenceof polynuclear aromatic hydrocarbon and2,3,7,8-tetrachloro-p-dioxin bindingproteins in the marine mussels, Bathymodiolus and Modiolus moliolus. AquaticToxicology,1999,48:51~64
[106] Witt G, Leipe T, Emeis K C. Using fluffy layer material to study the fate ofparticle-bound organic pollutants in the sourthern Baltic Sea. EnvironmentalScience&Technology,2001,35(8):1567~1573
[107] Witt G. Occurrence and transport of polycyclic aromatic hydrocarbons in thewater bodies of Baltic Sea. Marine Chemistry,2002,79(2):49~66
[108] Xu C Q, Pan L Q, Liu N, et al. Cloning, characterization and tissue distributionof a pi-class glutathione S-transferase from clam (Venerupis philippinarum):Response to Benzo[α]pyrene exposure. Comparative Biochemistry andPhysiology Part C,2010,152:160~166
[109] Zhang H, Song L S, Li C H, et al. Molecular cloning and characterization of athioester-containing protein from Zhikong scallop Chlamys farreri. MolecularImmunology,2007,44:3492~3500
[110] Zhang J F, Liu H, Sun Y Y, et al. Responses of the antioxidant defenses of theGoldfish Carassius auratus, exposed to2,4-dichlorophenol. EnvironmentalToxicology and Pharmacology,2005,19:185~190
[111] Zhang L, Li F G, Liu C L, et al. Construction and analysis of cotton (Gossypiumarboreum L.) drought-related cDNA library. BMC Research Notes,2009,2:120
[112] Zhang Z, Wang Y, Jiang Y, et al. Ribosomal protein124is differentiallyexpressed in ovary and testis of themarine shrimpmar-supenaeus japonicus.Biochem and Molecular Biology,2007,147(3):466~474
[113] Zhou J L, Maskaoui K. Distribution of polycyclic aromatic hydrocarbons inwater and surface sediments from Daya Bay, China. Environmental Pollution,2003,121(2):269~281
[114]安社娟,2005.多环芳烃致癌的分子毒理学研究进展.国外医学卫生学分册,32(1):10~13
[115]百度百科:《生物监测》,2012年4月24日,http://baike.baidu.com/view/469702.htm.
[116]百度百科:《抑制性消减杂交技术》,2010年12月7日,http://baike.baidu.com/view/4494504.htm.
[117]包艳波,徐日福,王志贤,等.鸡卵巢组织SSH cDNA文库构建及差异表达基因筛选与初步鉴定.吉林农业大学学报,2011.(http://www.cnki.net/kcms/detail/22.1100.s.20110516.1625.001.html)
[118]曾勇一,陆承平. cDNA微阵列结合抑制性差减杂交研究对虾白斑综合征病毒部分表达基因.病毒学报,2004,20(3):255~260
[119]陈亮,郭红岩,沈红,等.低浓度铅暴露对鲫鱼肝脏抗氧化系统的影响.环境化学,2002,21(5):485~489
[120]陈荣,刘辉,李东晓,等.水生动物谷胱甘肽硫转移酶研究进展.厦门大学学报自然科学版，2006,A02:176~184
[121]陈荣,郑微云,余群,等.石油污染对僧帽牡蛎(Ostrea cucullata)抗氧化酶的影响.环境科学学报,2002,22(3):385~388
[122]陈胜,董美阶,朱海燕,等.接触苯工人血浆热应激蛋白70抗体滴度分析.中华预防医学杂志,2000,34(1):8~10
[123]陈韬,苏建明,文祝友,章怀云,2003.扇贝性腺和肠组织表达序列标签的初步识别.农业生物技术学报.11(4):407~411
[124]单晓枫,孙佳敏,张洪波,等.抑制性消减杂交技术及其在水生动物基因克隆中的应用.黑龙江畜牧兽医,2010,12:30~32
[125]董瑞斌,许东风,刘雷,等.多环芳烃在环境中的行为.环境与开发,1999,14(2):10~11
[126]范志杰,宋春印.海洋环境污染生物监测及其方案的设计研究．交通环保，1994,1:3~8.
[127]方孝东,黄薇,林栖凤,等.通过cDNA RDA法分离和识别盐藻(Dunaliella salina)盐胁迫相关基因.中国生物化学与分子生物学报,2004,20(1):67~72
[128]韩菲,2007.多环芳烃来源与分布及迁移规律研究概述.气象与环境学报,23(4):57~61
[129]黄春,李润琴,张万江,2009.应用抑制性消减杂交技术筛选结核分枝杆菌差异基因.中华微生物学和免疫学杂志.6:507~512
[130]黄耀芬,李丽萍,胡显安,李再培.水质有机污染的指示生物——寡毛类.黑龙江环境通报,2003,2:71~72
[131]霍传林.近海环境中有机污染物对底栖鱼体内EROD活性的诱导作用研究:
[硕士学位论文].大连:大连理工大学.2003
[132]基因时代:《基因表达序列分析（SAGE）》,2009年9月14日,http://ggene.cn/html/protocol/bioinformatics/2009/0914/3126.html.
[133]吉成龙,孙国华,杨建敏,等.刺参(Apostichopus japonicus)高温胁迫消减cDNA文库的构建与分析.海洋与湖沼,2011,1:62~68
[134]江桂斌.持久性有毒污染物的环境化学行为与毒理效应.毒理学杂志,2005,19(3):179~180
[135]姜淑梅,吕晓梅,胡赞民.差减抑制杂交技术在植物研究中的应用.植物学通报,2005,22(增刊):91~98
[136]金赞芳,陈英旭.环境的PAHs污染及其生物修复技术研究进展.农业环境保护,2001,20(2):123~125
[137]孔繁祥.环境生物学.北京:高等教育出版社,2000.51-56
[138]匡少平,孙东亚.多环芳烃的毒理学特征与生物标记物研究.世界科技研究与发展,2007,29(2):41~47
[139]郎印海,贾永冈,刘宗峰,等.黄河口水中多环芳烃(PAHs)的季节分布特征及来源分析.中国海洋大学学报,2008,38(4):640~646
[140]李立芹,鲁黎明,2006.抑制性消减杂交技术（SSH）及其在烟草生物学研究中的应用.中国烟草科学.32(3):89~94
[141]李文雍,郁卫东,刘桂生,等.用DDRT—PCR技术克隆小鼠早期胚胎发育相关基因.中国生物化学与分子生物学报,2002, l8(6):728~732
[142]李小庆,景志忠.抑制性消减杂交技术的应用.生物技术通报,2009,5:46~50
[143]林道辉,朱利中,王静.小冶炼地区PAHs污染及其风险评价.生态学报,2005,25(2):262~267
[144]凌晰,刘文新,陈江麟,等.黄海近岸表层沉积物中PAHs的分布特征与潜在风险.环境科学学报,2008,28(7):1395~1399
[145]刘海芳,王凡.水环境污染程度的分子生物标志物的研究进展.安徽农业科学,2007,35(18):5502,5503~5506
[146]刘辉,叶湘漓.文昌鱼抗金葡菌感染差减文库的构建及免疫相关基因分析.中国生物化学与分子生物学报,2009,25(8):727~733
[147]刘晶,张文兵,麦康森,等.皱纹盘鲍外套膜耐维生素E缺乏消减cDNA文库的构建.中国水产科学,2007,14(3):383~389
[148]刘静.苯并a芘对栉孔扇贝血细胞免疫毒性机制的研究:[博士学位论文].青岛:中国海洋大学.2009
[149]刘玲.环芳烃的代谢和肺癌易感性的研究.国外医学卫生学分册,1986,3:136~140
[150]刘维全,高士争,王吉贵.分子生物学实验基本操作技术.北京:化学工业出版社,2010-04
[151]刘岩,张祖麟,洪华生.厦门西港表层海水PAHs含量分布特征和来源进行分析.海洋通报,1999,8(4):38~43
[152]刘移民.芳香烃类化合物受体研究进展.中国职业医学,2001,28(6):46~48
[153]马明辉,海志皆,冯志权,等.辽东湾双台子河海区动物体内污染物含量及时空分布.海洋环境科学,1999,18(1):61~76
[154]马伟业,千新来,赵清正,等.应用抑制性消减杂交技术分离E1A药敏相关基因.癌症,2004,23(2):146~149
[155]马长艳,周开亚,郭豫杰,等.中华绒螯蟹卵巢差减cDNA文库的构建.动物学研究,2003,24(1):53~56
[156]麦碧娴,林峥,张干,等.珠江三角洲沉积物中毒害有机物的污染现状及评价.环境科学研究,2001,14(1):19~23
[157]麦碧娴，林峥，张干,等.珠江三角洲河流和珠江口表层沉积物中有机污染物研究一多环芳烃和有机氯农药的分布及特征.环境科学学报,2000,2(2):192~197
[158]苗晶晶.苯并[a]芘对栉孔扇(Chlamys farreri)贝分子毒理学机制的研究:[博士学位论文].青岛:中国海洋大学,2010
[159]潘鲁青,刘娜,王静.栉孔扇贝在B[a]P胁迫下生物标志物筛选的研究.水生生物学报,2012,36(2):1~8
[160]庞朋沙,过倩萍,伍会健.细胞内AhR信号转导通路的机制研究.现代生物医学进展,2010,10(13):2567~2570
[161]戚元成,马雷,王菲菲,等.打顶烟草根尖组织抑制性消减cDNA文库的构建及其序列分析.中国农业科学,2011,44(7)
[162]丘耀文,周俊良, Maskaoui K.,等.大亚湾海域水体和沉积物中多环芳烃分布及其生态危害评价,2004,23(4):72~80
[163]曲跃军,杜人杰,王雷,等.抑制性消减杂交技术研究Cd2+胁迫下星星草基因表达.东北林业大学学报,2010,38(2):9~11
[164]饶志明,董海涛,庄杰云,等.水稻稻瘟病菌胁迫应答cDNA片段的表达及定位.中国水稻科学,2003,17(2):105~108
[165]任加云,潘鲁青,苗晶晶.苯并(a)芘和苯并(k)荧蒽混合物对栉孔扇贝毒理学指标的影响.环境科学学报,2006,26(7):1180~1186
[166]任天志.持续农业中的土壤生物指标研究.中国农业科学,2000,33(1):68~75
[167]任秀华,斯陆勤,曹磊,等.聚氧乙烯醚类表面活性剂对大鼠体内细胞色素P4503A活性的影响.药学学报,2008,43(5):528~534
[168]生物秀:《传统mRNA差异显示技术与第二代差异显示系统》,2005年11月30日, http://www.bbioo.com/experiment/12-926-1.html.
[169]苏建明.草鱼肠道组织消减cDNA文库的构建及免疫相关基因的克隆与表达分析:[博士学位论文].长沙:湖南农业大学,2007
[170]唐中林,万平,李小平,等.猪胚胎骨骼肌LongSAGE文库构建.农业生物技术学报,2007,15(1):171~172
[171]滕彦国,倪师军,张成江,等.攀枝花地区不同工业区表层土壤中重金属分布的特征.物探化探计算技术,2002,24(3):254~262
[172]田蕴,郑天凌,王新红.厦门西港表层沉积物中多环芳烃(PAHs)的含量、分布及来源.海洋与湖沼,2004,35(1):16~20
[173]田蕴,郑天凌,王新红.厦门西港表层海水中PAHs的含量、组成及来源.环境科学学报,2004,24(1):50~55
[174]王春光,李桂兰,尹松年.苯染毒小鼠体内DNA加合物及其持续时间的研究.中华预防医学杂志,1993,11:344~345
[175]王静,潘鲁青,苗晶晶,2007.多环芳烃化合物对栉孔扇贝组织芳烃羟化酶活力的影响.海洋科学.31(12):19~23
[176]王伟,2009. PLC法测定水中16种多环芳烃.环境科学与管理,34(7):108~111
[177]王晓蓉,罗义,施华宏,等.分子生物标志物在污染环境早期诊断和生态风险评价中的应用.环境化学,2006,25(3):320~325
[178]王秀丽,徐建民,姚槐应,等.重金属铜、锌、镉、铅复合污染对土壤环境微生物群落的影响.环境科学学报,2003,23(1):22~27
[179]王学彤,贾英,孙阳昭,等.典型污染区河流表层沉积物中PAHs的分布来源及生态风险.环境科学,2010,31(1):153~158
[180]王在照,焦传珍,张晓军,等.中国对虾蜕皮抑制激素全长cDNA的克隆及序列分析.遗传学报,2003,30(2):12~134
[181]王卓,张万起.芳香烃受体核转位蛋白的结构及相关功能.生命科学,2007,19(1):73~77
[182]徐凤山,张均龙.中国海典型生境双壳类软体动物多样性特点.生物多样性,2011,19(6):716~722
[183]许光辉,郑洪元.土壤微生物分析方法手册[M].上海:上海科技出版社,1986.226~228
[184]闫博,王兰,李涌泉,等.镉对长江华溪蟹肝胰腺抗氧化酶活力的影响.动物学报,2007,53(6):1121~1128
[185]阳晶.沙田柚花柱自交不亲和相关基因的克隆与鉴定:[硕士学位论文].桂林:广西师范大学,2008
[186]杨佰娟,郑立,陈军辉,等.南黄海中部表层沉积物中多环芳烃含量分布及来源分析,2009,29(3):663~667
[187]杨涛,陈海刚,蔡文贵,等.菲和苯并荧蒽曝露对翡翠贻贝外套膜的氧化胁迫及损伤.南方水产科学,2011,7(4):24~29
[188]杨毅,刘敏,侯立军,等.海岸带水环境中多环芳烃的归宿研究.海洋环境科学,2003,22(1):69~72
[189]杨志新,刘树庆.重金属Cd、Zn、Pb复合污染对土壤酶活性的影响.环境科学学报,2001,21(1):60~63
[190]叶细标,倪为民,傅华.分子生物标志物及其应用.中国工业医学杂志,2002,15(1):32~34
[191]于莹,张昌武,赵楠,等.胶州湾浮游纤毛虫丰度和生物量的周年变化.海洋与湖沼,2011,42(5):690~701
[192]袁东星,杨东宁,陈猛,等.厦门西港及闽江口表层沉积物中多环芳烃和有机氯污染物的含量及分布.环境科学学报，2001，21(1)：107~112
[193]岳桂东.玉米干旱胁迫相关基因的克隆与分析:[博士学位论文].济南:山东大学,2008
[194]岳敏,谷学新,邹洪.多环芳烃的危害与防治.首都师范大学学报,2003,24:40~44
[195]张娇.黄河及河口烃类有机物的分布特征及源解析:[博士学位论文].青岛:中国海洋大学,2008
[196]张强,张志文,龚侃,等.应用抑制性消减杂交技术克隆鉴定肾癌特异表达基因.科学通报,2000,45(16):1758~1762
[197]张先勇,刁晓平,杨宝，等.苯并[a]芘对马氏珠母贝血淋巴细胞DNA损伤的研究.生态环境学报,2011,20(6-7):1107~1110
[198]张新庆,杨佰娟,黎先春,等.南黄海中部海水中多环芳烃的分布特征.岩矿测试,2009,28(4):325~328
[199]张义兵,石耀华,桂建芳.鱼类培养细胞抗病毒基因差减cDNA文库的构建.水生生物学报,2003,27(2):113~118
[200]章卓,雷红宇,苏建明,2010.草鱼肠道组织抑制性消减表达序列标签的初步分析.湖南农业大学学报(自然科学版).36(3):339~343
[201]周驰,李纯厚,张为民，等.翡翠贻贝CYP4基因克隆及其表达水平分析.热带海洋学报,2010,29(4):82~88
[202]朱婧,杜立新:《反转录差异显示技术(DDRT-PCR)及其方法的改进》，2008年7月19日, http://www.biomart.cn/article/9/26/27/5993.htm.
[203]庄志雄,张桥.分子生物标志物研究的现状与展望.中华劳动卫生职业病杂志,1997,15(3):131~133
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