人参cDNA文库构建，EST与相关基因表达分析及EST-SSR标记建立
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摘要
人参为五加科多年生草本植物,是我国常用的珍贵药材。人参种质资源主要包括野生人参和栽培人参,野生人参为国家珍稀濒危物种。近年来,栽培人参品质下降、品种混乱、参地连作障害、生产滞销等原因严重破坏了人参种质资源。急需构建人参种质基因库,收集和整理人参宝贵的种质资源。人参作为珍贵的中药材资源,其体内次生代谢产物(人参皂苷等)具有多种生理功能,具有重要的药用价值。人参功能基因和次生代谢生物合成途径的研究相对较少,完全的基因组和草图序列还有待于建立。因此,保存人参种质资源和加速基因水平的研究进程具有重要意义。本文进行了人参叶片cDNA文库的构建,在此基础上进行了EST的初步分析,部分基因的时空表达分析,利用现有的人参EST资源进行SSR信息分析,人参EST-SSR标记的建立等方面的研究,结果如下:
通过四种RNA提取方法提取人参叶片总RNA,其中SDS法能分离出高质量的RNA,可以进行下一步的分子生物学研究;以四年生红果人参叶片为材料,提取叶片总RNA合成cDNA,连接到质粒载体pDNR-LIB上。采用电穿孔法将重组质粒转化到DH5a中,成功构建了人参叶片cDNA文库。经文库质量鉴定表明:原始文库滴度为1.008×10~6 pfu·mL~(-1),扩增后的文库滴度为2.968×10~9 pfu·mL~(-1),重组率接近100%,插入片段大小范围在0.5～2 kb之间,平均约为0.85 kb,表明应用SMART技术已成功构建了红果人参叶片cDNA文库,可供进一步的EST测序分析使用。对四年生红果人参叶片cDNA文库测序获得441条有效EST序列。GenBank登录号:ES554557-ES554997。441条EST拼接后代表了354个Unigenes。BLAST注释结果表明具有已知或推测功能基因136个,未知功能基因127个,未比对上的基因81个(可能是新基因)。根据BLASTX注释与GO分类结果,并参照MIPs分类标准对354个Unigenes按照生物学功能划分为9个大类:能量与代谢(energy/metabolism,13.8%)、蛋白合成(proteinsynthesis,5.9%)、信号传导(signal transduction,4.8%)、细胞救援与防御(cellrescue。defence,3.1%)、运输(transport,4.8%)、光合作用(photosynthesis,1.9%)、转录(transcription,2.2%)、蛋白活性调节(protein activity regulation,1.69%)、其他(others,61.5%)。
采用荧光定量RT-PCR技术研究人参叶、根部的部分基因的表达方式。结果表明,人参中的同一基因在叶、根部的表达模式不完全相同。所选择的基因的表达分别在人参皂苷生物合成途径、三羧酸循环途径、信号传导、抗逆、苯丙氨酸代谢途径等植物次生代谢中发挥重要作用。对人参基因时空表达的研究,将有利于我们进一步研究它们在人参代谢活动过程中的基因表达调控机理。
利用现有的7055条人参EST序列搜索出791个SSR,其出现频率为11.21%,平均长度为21.37 bp,平均分布频率为1/5.7 kb。人参EST-SSR的重复类型比较丰富,一至六核苷酸都能观测到。二核苷酸重复是主要的重复类型,占全部EST-SSR的56.89%,其次是三核苷酸重复的占全部SSR的21.11%。AT、GAA是二核苷酸和三核苷酸中出现次数最多的重复基元类型,分别占28.89%和10.18%。表明人参EST中的SSR数量非常丰富。本研究对现有人参EST资源中的SSR进行了分析,明确人参EST中SSR发生频率和分布特征信息,了解人参EST资源的特性,从而为建立其EST-SSR标记及其在人参分子标记的理论研究、人参遗传育种中的应用提供重要的依据。
根据搜索出的SSR设计了68对EST-SSR引物,对测试样品进行PCR扩增并检测多态性。分别以人参品种集安长脖和抚松二马牙的DNA为模板,对设计的EST-SSR引物进行筛选,发现有43对EST-SSR引物能扩增出产物。在9个人参品种、2个西洋参品种和2个刺五加品种中进一步对这些可扩增的引物对进行多态性检测,发现有26对引物显示多态性,占可扩增引物的60.47%,占设计引物总数的38.23%。表明实验建立的EST-SSR标记是一种行而有效的方法,是对现有人参EST资源的进一步开发和利用。建立这种标记对于加速人参EST资源的开发利用、丰富其分子标记类型与绘制遗传图谱、实现特定性状的辅助选择和遗传多样性等研究都具有重要的意义。目前人参中还没有建立EST-SSR标记的报道,本研究根据现有人参EST资源首次建立了EST-SSR标记。
Ginseng(Panax ginseng C.A.Meyer),a perennial herb from the Araliaceae family,is one of the most commonly utilized medicinal plants.Ginseng germplasm resources mostly involve in wild ginseng and cultural ginseng.The wild ginseng is considered as rare endangered species in China.At present,Ginseng germplasm resources are destroyed because of quality dropped, variety confusion,injury by continuous cropping,produce dull sale.ect.So we need to construct the gene library of ginseng germplasm,collecte and rearrange the precious germplasm resources.Ginseng as rare traditional Chinese medicinal materials,their secondary metabolite(ginsengoside etc.) in vivo are important effective constituents in biological activity of ginseng and possess important medicinal value.The research about functional gene and secondary metabolism biosynthetic pathway is relatively less in ginseng.In case of Panax ginseng,Hawerever,the full genome and draft sequences are yet to be established.It is meaningful for preserving panax ginseng germplasm resources and accelerating to studying the gene level process.In this paper,we reported the construction of the cDNA library from panax ginseng leaves,EST analysis,space-time expression analysis of gene,analysis of SSR of EST resources in panax ginseng,development of EST-SSR marker in panax ginseng.The main results are as follows.
RNA was isolated from panax ginseng leaves using four methods of extraction(CTAB, Trizol,Bizol,SDS).Highly-grade RNA was isolate by using SDS method,better than the other methods and suitable to the demands of molecular biological research.The four years old red fruit panax ginseng was used as experimental material,and the total RNA was extract from the leaves,the cDNA was synthesized and the ds cDNA fragment was ligated into the pDNR-LIB vector.The recombinant plasmid was transformed into E.coli DH5 a by electroporation.The library qualification evaluation showed:the titer of primary cDNA library was 1.008×106 pfu·mL~(-1),the titer of amplified library was 2.968×109 pfu·mL~(-1),the percentage of recombination was about 100%,the fragment size of inserted was 0.5～2kb.The average insert size was 0.85 kb.The cDNA library was constructed successfully,and could be satisfied for further studying on expressed sequence tags(ESTs) analysis.A total of 441 high quality ESTs were obtained from cDNA library of panax ginseng leaves.The GenBank accession numbers are ES554557-ES554997.The 441 ESTs were assembled into 354 unigenes.The results of BLAST annotation showed:36 known or putative functional gene,127 unknown gene,81 no hits(maybe new genes).The 354 unigenes were divided into 9 functional categorier based on BLASTX,GO analysis and MIPs Functional Catalogue,i.e.,energy/metabolism(13.8%), protein synthesis(5.9%),signal transduction(4.8%),cell rescue,defence(3.1%), transport(4.8%),photosynthesis(1.9%),transcription(2.2%),protein activity regulation (1.69%),others(61.5%0).
To studying gene expressed mode between roots and leaves of panax ginseng in different growth time,eight genes from panax ginseng were selected for expression analysis using real time revers transxription-polymerase chain reaction(RT-PCR).The results showed the expressed mode of the same gene was different or incomplete the same.Eight genes studied played the important roles in ginsenoside biosynthetic pathway,tricarboxylic cycle,signal conduct,stress-tolerance,phenylalanine metabolic pathway and so on.To study gene spacetime expressed in panax ginseng,there will be benefit to further study regulation and control mechanism of gene expression in metabolic activity.
Seven hundred and ninety one microsatellites(SSRs) were isolated from 7055 panax ginseng expressed sequence tags(ESTs).The occurrence frequency of SSR was 11.21%,the average length searched was 21.37 bp and the average distance of distribution was 1/5.7 bp. The dinucleotide repeat was the dominant repeat type in panax ginseng EST-SSR,accounting for 56.89%of the total EST-SSR,and then trinucleotide repeat accounts for 21.11%of the total EST-SSR.AT and GAA were the most frequent motifs,accounting for 28.89%and 10.18%(?) dinucleotide and trinucleotide repeats,respectively.SSR quantities were abundant in panax ginseng ESTs,SSR occurrence frequency and distributional characteristixs of EST resources were definited,which will provide important references for development of EST-SSR marker and application in inherit breeding of panax ginseng.
According to primer design criteria,sixty-eight primer pairs for EST-SSR were designed. Under an appropriate PCR reaction system,all EST-SSR primer pairs were screened ag(?)inst genomic DNA of ji'anchangbo and fusong'ermaya from panax ginseng respectively,and fortythree of sixty-eight EST-SSR primer pairs resulted in PCR products.Then all forty-three EST-SSR primer pairs available were detected in nine Panax ginseng cultivars,two Panax quinquefolius cultivars and two Acanthopanax senticosus cultivars for polymorphisms,and twenty-six EST-SSR primer pairs were polymorphic,accounting for 60.47%of primer p(?)s amplified,accounting for 38.23%of total primer pairs designed.These results showed that it is an effective and feasible approach to develop EST-SSR markers using Panax ginseng ESTs. Developing of ginseng EST-SSR markers will further detect and make full use of ginseng ESTs resources.It is meaningful for accelerating to exploiting and utilizing of the ginseng ESTs resource,enriching molecular marker types,constructing genetic map,assessing genetic diversity,and assisting plant breeding.At present,there is no report about developing of ginseng EST-SSR marker in domestic and foreign.
通过四种RNA提取方法提取人参叶片总RNA,其中SDS法能分离出高质量的RNA,可以进行下一步的分子生物学研究;以四年生红果人参叶片为材料,提取叶片总RNA合成cDNA,连接到质粒载体pDNR-LIB上。采用电穿孔法将重组质粒转化到DH5a中,成功构建了人参叶片cDNA文库。经文库质量鉴定表明:原始文库滴度为1.008×10~6 pfu·mL~(-1),扩增后的文库滴度为2.968×10~9 pfu·mL~(-1),重组率接近100%,插入片段大小范围在0.5～2 kb之间,平均约为0.85 kb,表明应用SMART技术已成功构建了红果人参叶片cDNA文库,可供进一步的EST测序分析使用。对四年生红果人参叶片cDNA文库测序获得441条有效EST序列。GenBank登录号:ES554557-ES554997。441条EST拼接后代表了354个Unigenes。BLAST注释结果表明具有已知或推测功能基因136个,未知功能基因127个,未比对上的基因81个(可能是新基因)。根据BLASTX注释与GO分类结果,并参照MIPs分类标准对354个Unigenes按照生物学功能划分为9个大类:能量与代谢(energy/metabolism,13.8%)、蛋白合成(proteinsynthesis,5.9%)、信号传导(signal transduction,4.8%)、细胞救援与防御(cellrescue。defence,3.1%)、运输(transport,4.8%)、光合作用(photosynthesis,1.9%)、转录(transcription,2.2%)、蛋白活性调节(protein activity regulation,1.69%)、其他(others,61.5%)。
采用荧光定量RT-PCR技术研究人参叶、根部的部分基因的表达方式。结果表明,人参中的同一基因在叶、根部的表达模式不完全相同。所选择的基因的表达分别在人参皂苷生物合成途径、三羧酸循环途径、信号传导、抗逆、苯丙氨酸代谢途径等植物次生代谢中发挥重要作用。对人参基因时空表达的研究,将有利于我们进一步研究它们在人参代谢活动过程中的基因表达调控机理。
利用现有的7055条人参EST序列搜索出791个SSR,其出现频率为11.21%,平均长度为21.37 bp,平均分布频率为1/5.7 kb。人参EST-SSR的重复类型比较丰富,一至六核苷酸都能观测到。二核苷酸重复是主要的重复类型,占全部EST-SSR的56.89%,其次是三核苷酸重复的占全部SSR的21.11%。AT、GAA是二核苷酸和三核苷酸中出现次数最多的重复基元类型,分别占28.89%和10.18%。表明人参EST中的SSR数量非常丰富。本研究对现有人参EST资源中的SSR进行了分析,明确人参EST中SSR发生频率和分布特征信息,了解人参EST资源的特性,从而为建立其EST-SSR标记及其在人参分子标记的理论研究、人参遗传育种中的应用提供重要的依据。
根据搜索出的SSR设计了68对EST-SSR引物,对测试样品进行PCR扩增并检测多态性。分别以人参品种集安长脖和抚松二马牙的DNA为模板,对设计的EST-SSR引物进行筛选,发现有43对EST-SSR引物能扩增出产物。在9个人参品种、2个西洋参品种和2个刺五加品种中进一步对这些可扩增的引物对进行多态性检测,发现有26对引物显示多态性,占可扩增引物的60.47%,占设计引物总数的38.23%。表明实验建立的EST-SSR标记是一种行而有效的方法,是对现有人参EST资源的进一步开发和利用。建立这种标记对于加速人参EST资源的开发利用、丰富其分子标记类型与绘制遗传图谱、实现特定性状的辅助选择和遗传多样性等研究都具有重要的意义。目前人参中还没有建立EST-SSR标记的报道,本研究根据现有人参EST资源首次建立了EST-SSR标记。
Ginseng(Panax ginseng C.A.Meyer),a perennial herb from the Araliaceae family,is one of the most commonly utilized medicinal plants.Ginseng germplasm resources mostly involve in wild ginseng and cultural ginseng.The wild ginseng is considered as rare endangered species in China.At present,Ginseng germplasm resources are destroyed because of quality dropped, variety confusion,injury by continuous cropping,produce dull sale.ect.So we need to construct the gene library of ginseng germplasm,collecte and rearrange the precious germplasm resources.Ginseng as rare traditional Chinese medicinal materials,their secondary metabolite(ginsengoside etc.) in vivo are important effective constituents in biological activity of ginseng and possess important medicinal value.The research about functional gene and secondary metabolism biosynthetic pathway is relatively less in ginseng.In case of Panax ginseng,Hawerever,the full genome and draft sequences are yet to be established.It is meaningful for preserving panax ginseng germplasm resources and accelerating to studying the gene level process.In this paper,we reported the construction of the cDNA library from panax ginseng leaves,EST analysis,space-time expression analysis of gene,analysis of SSR of EST resources in panax ginseng,development of EST-SSR marker in panax ginseng.The main results are as follows.
RNA was isolated from panax ginseng leaves using four methods of extraction(CTAB, Trizol,Bizol,SDS).Highly-grade RNA was isolate by using SDS method,better than the other methods and suitable to the demands of molecular biological research.The four years old red fruit panax ginseng was used as experimental material,and the total RNA was extract from the leaves,the cDNA was synthesized and the ds cDNA fragment was ligated into the pDNR-LIB vector.The recombinant plasmid was transformed into E.coli DH5 a by electroporation.The library qualification evaluation showed:the titer of primary cDNA library was 1.008×106 pfu·mL~(-1),the titer of amplified library was 2.968×109 pfu·mL~(-1),the percentage of recombination was about 100%,the fragment size of inserted was 0.5～2kb.The average insert size was 0.85 kb.The cDNA library was constructed successfully,and could be satisfied for further studying on expressed sequence tags(ESTs) analysis.A total of 441 high quality ESTs were obtained from cDNA library of panax ginseng leaves.The GenBank accession numbers are ES554557-ES554997.The 441 ESTs were assembled into 354 unigenes.The results of BLAST annotation showed:36 known or putative functional gene,127 unknown gene,81 no hits(maybe new genes).The 354 unigenes were divided into 9 functional categorier based on BLASTX,GO analysis and MIPs Functional Catalogue,i.e.,energy/metabolism(13.8%), protein synthesis(5.9%),signal transduction(4.8%),cell rescue,defence(3.1%), transport(4.8%),photosynthesis(1.9%),transcription(2.2%),protein activity regulation (1.69%),others(61.5%0).
To studying gene expressed mode between roots and leaves of panax ginseng in different growth time,eight genes from panax ginseng were selected for expression analysis using real time revers transxription-polymerase chain reaction(RT-PCR).The results showed the expressed mode of the same gene was different or incomplete the same.Eight genes studied played the important roles in ginsenoside biosynthetic pathway,tricarboxylic cycle,signal conduct,stress-tolerance,phenylalanine metabolic pathway and so on.To study gene spacetime expressed in panax ginseng,there will be benefit to further study regulation and control mechanism of gene expression in metabolic activity.
Seven hundred and ninety one microsatellites(SSRs) were isolated from 7055 panax ginseng expressed sequence tags(ESTs).The occurrence frequency of SSR was 11.21%,the average length searched was 21.37 bp and the average distance of distribution was 1/5.7 bp. The dinucleotide repeat was the dominant repeat type in panax ginseng EST-SSR,accounting for 56.89%of the total EST-SSR,and then trinucleotide repeat accounts for 21.11%of the total EST-SSR.AT and GAA were the most frequent motifs,accounting for 28.89%and 10.18%(?) dinucleotide and trinucleotide repeats,respectively.SSR quantities were abundant in panax ginseng ESTs,SSR occurrence frequency and distributional characteristixs of EST resources were definited,which will provide important references for development of EST-SSR marker and application in inherit breeding of panax ginseng.
According to primer design criteria,sixty-eight primer pairs for EST-SSR were designed. Under an appropriate PCR reaction system,all EST-SSR primer pairs were screened ag(?)inst genomic DNA of ji'anchangbo and fusong'ermaya from panax ginseng respectively,and fortythree of sixty-eight EST-SSR primer pairs resulted in PCR products.Then all forty-three EST-SSR primer pairs available were detected in nine Panax ginseng cultivars,two Panax quinquefolius cultivars and two Acanthopanax senticosus cultivars for polymorphisms,and twenty-six EST-SSR primer pairs were polymorphic,accounting for 60.47%of primer p(?)s amplified,accounting for 38.23%of total primer pairs designed.These results showed that it is an effective and feasible approach to develop EST-SSR markers using Panax ginseng ESTs. Developing of ginseng EST-SSR markers will further detect and make full use of ginseng ESTs resources.It is meaningful for accelerating to exploiting and utilizing of the ginseng ESTs resource,enriching molecular marker types,constructing genetic map,assessing genetic diversity,and assisting plant breeding.At present,there is no report about developing of ginseng EST-SSR marker in domestic and foreign.
引文
[1]鱼红闪,陈琪,金凤燮.不同种类人参及其各部位中皂苷组成和比例的研究.食品与发酵工业,2002,28(2):24-28
[2]窦德强,靳玲,陈英杰.人参的化学成分及药理活性的研究进展与展望.沈阳药科大学学报,1999,16(2):151-156
[3]张雅玉.人参皂苷分析方法的研究进展,特产研究,1998,1:50-52
[4]章观德.人参的化学成分和分析方法研究进展,药学学报,1980,15(6):375-384
[5]Ma YC,Luo M,Mally L,Docet M.Distribution and proportion of major ginsenosides and qualty control of ginseng products.Chinese Journal of medical chemistry,1996,6:11-21
[6]柳鹏,李勇.人参皂甙生物活性的研究进展.中国生育健康杂志,2004,15(5):317-319
[7]别光增道.药用人参及其有关植物成分的研究-西洋参的微量皂苷及所谓巴西人参的成分.国外医学中医中药分册,1983,4:42-45
[8]颜春芳,马家锐.人参植株不同部位皂甙含量研究.现代应用药学,1991,8(1):47-48
[9]徐绥绪,陈英杰,吕永俊,王志学,王乃利,姚新生.人参和国产西洋参化学的系统研究.沈阳药学学报,1989,6(4):286-289
[10]郑友兰,张崇禧,李向高.人参和西洋参芦头与根的化学成分比较分析.吉林农业大学人参科技论文集,1988,8:433-437
[11]张翼伸,李润秋,王玉万.人参多糖的研究.东北师范大学学报(Ⅰ),1982,2:97-103
[12]梁忠岩,张翼伸,苗春艳.西洋参与人参总糖组成及含量比较研究.中国药学杂志,1989,24(4):207-209
[13]Ji-Hye Lee and Kyung Hyun Kim.Pectin-like acidic polysaccharides from Panax ginseng with selective antiadhesive activity against pathogenic bacteria.Carbohydrate search,2006,341:1154-1163
[14]沈宁,王素贤,吴立军,李铣.中国吉林栽培西洋参挥发油的新近研究.沈阳药学院学报,1991,8(3):175-181
[151郑友兰,张崇禧,李向高.人参和西洋参芦头与根的化学成分比较分析.吉林农业大学.人参科研论文集,1988.8:433-436
[16]王勇,石磊,王慧,孙允秀,刘在群,罗旭阳,王建辉,林英杰,洪哲.人参茎叶挥发油中倍半萜烯类化合物的分离与鉴定.吉林大学自然科学学报,2001,1:88-90
[17]唐斌,程绪菊,刘江,唐冰.人参皂苷药理作用研究进展.西南军医,2005,7(3):45-47
[18]姜正林,吴新民,陈云,金淑仪,陈燕萍,庄坚.人参皂甙的抗鼠脑缺氧损伤作用.中华航海医学杂志,2000,7(1):28-32
[19]Kim YC,Kim SR,Markelonis GJ,Oh TH.Ginsenosides Rb1and Rg3 protect cultured rat cortical cells from glutamate-induced neurodegeneration.J Neurosci Res,1998,53:426-432
[20]Radad K,Gille G,Moldzio R,Ishige K,Rausch WD.Ginsenosides Rb1 and Rgl effects on survival and neurite growth of MPP(+)-affected mesencephalic dopaminergic cells.J Neural Transm,2004,111:37-45
[21]候明晓,敖定椿.人参总皂甙抗心肌缺血-再灌注损伤的作用机制.中国胸心血管外科临床杂志,2000,7(4):256-259
[22]Okamura N,Kobayashi K,Akaike A,Yagi A.Protective effect of ginseng saponins against impaired brain growth in neonatal rats exposed to ethanol.Biol Pharm Bull,1994,17:270-274
[23]白长喜.人参皂甙 Re 对大鼠心肌缺血-在灌注损伤时心肌和血浆前列腺素 E 水平的影响.延边医学院学报,1993,16(1):17-19
[24]张南生,张秀华,李文峰.人参皂苷 Rg3研究进展.医药导报,2006,25(7):687-689
[25]Mochizuki M,Yoo Yc,Matsuzawa K,Satok K,Saiki I,Tono-oka S,Samukawa K,Azuma I.Inhibitory effect of tumor metastasis in mice by saponins,ginsenoside Rb2,20(R)-and 20(S)-ginsenoside-Rg3,of red ginseng.Biol Pharm Bull,1995,18:197-1202
[26]高勇,王杰军,许青,叶琴琴,郭静,耿怀成.人参皂甙 Rg3抑制肿瘤心神血管形成的研究.第二军医大学学报,2001,22(1):40-42
[27]Nakata H,Kikuchi Y,Tode T,Hirata J,Kita T,Ishii K,Kudoh K,Nagata I,Shinomiya N.Inhibitory effects of ginsenoside Rh2 on tumor growth in nude mice bearing human ovarian cancer cells.Japanese Journal of Cancer Research,1998,89:733-740.
[28]王巍,杜松洁,王乃功.人参皂甙 Rb1和 Rg1对幼小鼠性腺和副性腺的影响.中药药理与临床,1999,15(1):10-11
[29]王艳华,肖雪媛,杨贵贞.人参皂甙对手术应激大鼠下丘脑-垂体-肾上腺轴及免疫靶器官中肾上腺皮质激素释放、ACTH 阳性细胞数的影响.中国神经免疫学和神经病学杂志,1998,5(1):12-15
[30]Haralampidis K,Trojanowska M,Osbourn AE.Biosynthesis of triterpenoid saplonins in plants.Adv Biochem Eng Biotechnol,2002,75:31-49
[31]Walker K,Fuj isaki S,Long R,Croteau R.Molecular cloning and heterologous expression of the C-13 phenylpropanoid side chain-CoA acyltransferase that functions in Taxol biosynthesis.Proc Natl Acad Sci USA,2002,99:12715-12720
[32]Wallaart TE,Bouwmeester HJ,Hille J,Poppinga L,Maijers NC.Amorpha-4,11-diene synthase:Cloning and functional expression of a key enzyme in the biosynthetic pathway of the novel antimalarial drug artemisinin.Planta,2001,212:460-465
[33]Yazaki K,Kunihisa M,Fujisaki T,Sato F.Geranyl diphosphate:4-Hydroxybenzoate geranyltransferase from Lithospermum erythrorhizon.Cloning and characterization of a key enzyme in shikonin biosynthesis.J Biol Chem,2002,277:6240-6246
[34]Grothe T,Lenz R,Kutchan TM.Molecular characterization of the salutaridinol 7-O- acetyltransferase involved in morphine biosynthesis in opium poppy Papaver somniferum.J Biol Chem,2001,276:30717-30723
[35]赵寿经,李昌禹,臧埔,马海琴,杨振堂.影响人参发根中皂甙含量基因的定位及其克隆.特产研究,2001,4:1-5
[36]Bulgakov VP,Khodakovskaya MV,Labetskaya NV,Tchernoded GK,Zhuraclev YN.The impact of plant rolC oncogene on ginsenoside production by ginseng hairy root cultures.Phytochemistry,1998,49:1929-1934
[37]罗志勇,陆秋恒,刘水平,陈湘晖,罗建清,汤立军,胡维新.人参植物皂苷生物合成相关基因的筛选与鉴定.生物化学与生物物理学,2003,35(6):554-560
[38]刘水平,罗志勇,陈湘晖,文斌,汪凌昊,魏来,胡维新.人参皂苷生物合成相关新基因GBR6的 cDNA 克隆及序列分析.生命科学研究,2004,8(4):351-354
[39]王昆,王颖,鲍永利孟祥颖,乌垠,易静文,李玉新.人参叶全长 cDNA 文库的构建及部分克隆的序列分析.分子科学学报,2006,22(1)58-62
[40]胡向阳,张文清,方建颖,蔡伟明,汤章城.脱乙酰壳多糖处理增加人参细胞皂苷的累积和皂苷合成关键酶基因的转录.植物生理与分子生物学学报,2002,28(6):485-490
[41]胡向阳,方建颖,蔡伟明,汤章城.模拟失重对钙调蛋白基因表达与人参皂苷合成的影响.自然科学进展,2005,5(13):537-540
[42]Jung,JD,Park HW,HahnY,Hur CG,In DS,Chung H J,Liu JR,Choi DW.Discovery of genes for ginsenoside biosynthesis by analysis of ginseng expressed sequence tags.Plant Cell Rep,2003,22:224-230
[43]Kim MK,Lee BS,In JG,Sun H,Yoon JH,Yang DK.Comparative analysis of expressed sequence tags(ESTs) of ginseng leaf.Plant Cell Rep,2005,25:599-606
[44]Choi D W,Jung JD,Ha YI,Park HW,In DS,Chung HJ Liu JR.Analysis of transcripts in methyl jasmonate-treated ginseng hairy roots to identify genes involved in the biosynthesis of ginsenosides and other secondary metabolites Plant Cell Rep,2005,23:557-566
[45]Galaud JP,Carri(?)re M,Pauly N,Canut H,Chalon P,Caput D,Pont-ezica RF.Construction of two ordered cDNA libraries enriched in genes encoding plasmalemma and tonoplast proteins from a high-efficiency expression library.Plant J,1999,17:111-118.
[46]Hawkins V,Doll D,Bumgarner R,Smith T,Abajian C,HooD L,Nelson PS.PEDB:the Prostate Expression Database.Nucleic Acids Res,1999,27:204-208
[47]Peterson LA,Brown MR,Carlisle A J,Kohn EC,Liotta LA,Emmert-Buck MR,Krizman DB.An improved method for construction of directionally cloned cDNA libraries from microdissected cells.Cancer Res,1998,58:5326-5328
[48]Diatchenko L,Lau YF,Campbeu AP,Chenchik A,Moqadam F,Huang B,Lukyanov S,Lukyanov K,Gurskaya N,Sverdlov ED,Siebert PD.Suppression subtractive hybridization:a method for generating differentially regulated or tissue-specific cDNA probes and libraries.Proc Natl Acad Sci USA,1996,93:6025-6030
[49]王关林,方宏筠．植物基因工程(第二版)．北京：科学技术出版社,2002：43～170
[50]Young RA and Davis RW.Efficient isolation of genes by using antibody probes.Proc Natl Acad Sci USA,1983,80:1194～1198
[51]Aguan K,Kusano T,Suzuki N,Kitagawa Y.An improved method for the construction of high efficiency cDNA library in plasmid or lambda vector.Nucleic Acids Res,1990,8:1071～1076
[52]Gonos ES,Derventzi A,Kveiborg M,Agiostratidou G,Kassem M,Clar BFC,Jat PS,Rattan SIS.Cloning and identification of genes that associated with mammalian replicative senescence.Exp Cell Res,1998,240:66～74.
[53]Hakvoort TB,Spijkers JA,Vermeulen JL,Lamers WH.Preparation of a differentially expressed full-length cDNA expression library by RecA-mediated triple-strand formation with subtractively enriched cDNA fragments.Nucleic Acids Res,1996,24:3478～3480.
[54]Gurskaya NG,Diatchenko L,Chenchik A,Siebert PD,Khaspekov GL,Lukyanov KA,Vagner LL,Ermolaeva OD,Lukyanov SA,Sverdlov ED.Equalizing cDNA subtraction based on selective suppression of Polymerase Chain Reaction:cloning of Jurkat transcripts induced by phytohemaglutinin and phorbol 12-myristable 13-acetable.Analytical Biochemistry,1996,240:90～97
[55]Wong BR,Rho J,Arron J,Robinson E,Orlinick J,Chao M,Kalachikov S,Cayani E,Bartlett FS 3rd,Frankel WN,Lee SY,Choi Y.Trance is a novel ligand of the tumor necrosis factor receptor family that activates c-Jun N-terminal kinase in T cells.J Biol Chem,1997,272:25190～25194
[56]Chu,ZL,McKinsey TA,Liu L,Gentry JJ,Malim MH,Ballard DW.Suppression of tumour necrosis factor-induced cell death by inhibitor of apoptosis C-IAP2 is under NF-B control.Proc Natl Acad Sci USA,1997,94:10057～10062
[57]Siki RK,Scharf S,Faloona F,Mullis KB,Horn GT,Erlich HA,Arnheim.Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.Science.1985,230:1350～1354
[58]Belyavsky A,Vinogradova T,and Rajewsky K.PCR-based cDNA library construction:general cDNA libraries at the level of a few cells.Nucleic Acids Res,1989,17:2919～2932.
[59]Fromont_Racine M.,Bertrand E,Pictet R,Grange T.A highly sensitive method for mapping the 5' termini of mRNAs.Nucleic Acids Res,1993,21:1683～1684
[60]Suzuki Y,Yoshitomo-Nakagawa K,Maruyama K,Suyama A,Sugano S.Construction and characterization of a full length-enriched and a 5'-end-enriched cDNA library.Gene,1997,200:149～156
[61]Frohman MA,Dush MK,and Martin G.R.Rapid production of full-length cDNAs from rare transcripts:Amplification using a single gene-specific oligonucleotide primer.Proc Natl Acad Sci USA,1988,85:8998-9002
[62]Roeder T.Solid-phase cDNA library construction,a versatile approach.Nucleic Acids Res,1998,26:3451-3452
[63]Rondinelli RH,Tricoli JV.CLARl,a novel gene that exhibits enhanced expression in advanced human prostate cancer.Clin Cancer Res,1999,5:1595-1602
[64]Velculescu VE,Zhang L,Vogelstein B,Kinzler KW.Serial analysis of gene expression.Science,1995,270:484-487
[65]Sompayrac L,Jane S,Bum TC,Tenen DG,and Danna KJ.Overcoming limitations of the mRNA differential display technique.Nucleic Acids Res,1995,23:4738-4739
[66]Adams MD,Kelley JM,Gocayne JD,Dubnick M,Polymeropoulos MH,Xiao H,Merril CR,Wu A,Olde B,Moreno RF.Complementary DNA sequencing:Expressed Sequence Tags and Human Genome Project.Science,1991,252:1651-1656
[67]White JA,Todd J,Newman T,Focks N,Girke T,de Il(?)rduya OM,Jaworski JG,Ohlrogge JB,Benning C.A New set of Arabidopsis Expressed Sequence Tags from Developing Seeds.The etabolic Pathway from Carbohydrates to Seed Oil.Plant Physiol,2000,124:1582-1594
[68]Bouchez D and Hofte H.Functional genomic in plants.Plant Physiology.1998,118:725-732
[69]Ewing RM,Kahla AB,Poirot O.Large-scales statistical analysis-of rice ESTs reveal correlate Kahlad patterns of gene expression.Genome Res,1999,9:950-959
[70]Adams MD,Dubnick M,Kerlavage AR,Moreno R,Kelley JM,Utterback TR,Nagle JW,Fields C,Venter JC.Sequence identification of 2,375 human brain genes.Nature,1992,355:632-634
[71]Asamizu E,Nakamura Y,Sato S,Tabata S.Characteristics of the Lotus japonicus gene repertoire deduced from large-scale expressed sequence tag(EST) analysis.Plant Mol Biol,2004,54:405-414
[72]Wan JS,Sharp S J,Poirier G.M.Cloning differentially expressed mRNA.Nature Biotechnol,1996,14:1685-1691
[73]李靖,韩本立,黄桂君,钱桂生,徐晓霞,杨彤翰,梁平,陈杰.应用抑制消减杂交技术构建肝癌差异表达基因消减 cDNA 文库.第三军医大学学报,2001,23(6):698-701
[74]罗志勇,刘水平,陆秋恒,陈湘晖,文斌,罗建清,胡维新.人参植物与皂苷生物合成相关的差减 cDNA 文库构建及基因差异表达分析.生命科学研究.2003,71(4):324-328
[75]李明芳,郑学勤.开发 SSR 引物方法之研究动态.遗传.2004,26(5):769-776
[76]曾庆国,陈艺燕.微卫星位点筛选方法综述.生态科学,2005,24(4):368-372
[77]Schena M,Schalon D,Davis RW and Brown PQ.Quantitative monito ring of gene expression patterns with a complementary DNA microarray.Science,1995,270:467-470
[78]Schena M.Genome analysis with gene expression microarrays.Bioassays,1996,18:427-431
[79]杨传平,王玉成,刘桂丰,姜静,张国栋.基因芯片技术研究柽柳 NaHCO3 胁迫下基因的表达.生物工程学报,2005,21(2):220-226
[80]祝骥,马文丽,姚汝华.DNA 芯片技术与基因表达研究.生命科学研究.2000,4(2):106-111
[81]Hatey F,Tosser-Klopp G,Clouscard-martinato C,Mulsant P and Gasser F.Expressed sequenced tags for genes:a review.Genet Sel Evol,1998,30:521-541
[82]Zhou Y,Tang JB,Walker MG,Zhang XQ,Wang J,Hu SN.Gene identification and expression analysis of 86136 sequencetags(EST) from the rice genome.Gen Prot Bioinfo,2003,1:26-42
[83]Rossi M,Araujo PG,Paulet F,Garsmeur O,Dias VM,Chen H,Van Sluys MA,D Hont A.Genomic distribution and characterization of EST-derived resistance gene analogs(RGAs)in sugarcane.Mol Gen Genomics,2003,269:406-419
[84]Rounsley S,Linx KK.Large scale sequencing of plant genome.Curr Opin Plant Biol,1998,1:136-141
[85]Sasaki T.The rice genome project in Japan.Proc Natl Acad Sci USA,1998,95:2027-2028
[86]Kurata N,Nagamura Y,Yamamoto K,Sue N,Wu J,Antonio BA,Shomura A,Shimizu T,Lin SY,Inoue T,Fukuda A,Shimano T.Nagamura Y,Yamamoto K,et al.A 300 kilobase interval genetic map of rice including 883 expressed sequences.Nat Genet,1994,8:365-372
[87]Chee PW,Rong J,William-Coplin D.EST derived PCR-based markers for functional gene homologues in cotton.Gene,2004,47:449-462
[88]Kotani H,Nakamura Y,Sato S,Kaneko T,Asamizu E.Structural analysis of Arabidopsis thaliana chromosome 5.VL.Sequence features of the regions of 1,367,185bp covered by 19physically assigned P1 and TAC clones.DNA Res,1998,5:203-216
[89]Schuler GD,Boguski MS,Stewart EA,Stein LD,Gyapay G,Rice K,White RE.ect.A gene map of the human genome.Science,1996,274:540-546
[90]Cui YL,Zhang XY,Zhou Y,Yu H,Tao L,Zhou J,Zhuge Q,Cai YM,Huang MR..Identification and expression Analysis of EST-based Genes in the Bud of Lycoris longituba.Geno Prot Bioinfo,2004,2:42-46
[91]Shin KE,Mary AC,Inna A,Davina B,Monica F,Elizabeth C,John CC.Transcript profiling of salinity stress responses by large-scale expressed sequence tag analysis in Mesembryanthemum crystallinum.Gene,2004,341:83-92
[92]王华春,陈清轩.充分利用 EST 数据库资源.生物化学与生物物理进展,2000,27:442-444
[93]Chen X,Salamini R and Gebhardt C.A Potato molecularfunction map for carbohydrate metabolism and transport.Theor Appl Genet,2001,102:284-295
[94]Thiel T,Miehalek W,Varshney RK,Graner A.Exploiting EST databases for the development and characterization of gen-derived SSR-markers in barley(Hordeum vulgareL.).Theor Appl Genet,2003,106:411-422
[95]Holton RA,Christopher JT,McClure L,Harker N,Henry RJ.Identification and mapping of polymorphic SSR markers from expressed gene sequences of barley and wheat.Mol Breeding,2002,9:63-71
[96]陈亮,赵丽萍,高其康.茶树新梢 cDNA 克隆测序和表达序列标签(ESTs)特性分析.农业生物技术学报,2005,13(1):21-25
[97]王昆,王颖,鲍永利,孟祥颖,乌垠,李玉新.提取高质量人参 RNA 的方法研究.生物化学与生物物理进展,2006,33(1):95-99
[98]王壮伟,渠慎春,章镇,张君毅.苹果属 RNA 高效快速提取新方法.果树学报,2004,21(4):385-387
[99]Wang DH,Wang BC,Li B,Duan CR,Zhang J.Extraction of total RNA from Chrysanthemum containing high levels of phenolic and carbohydrates.Colloid Surface B,2004,36:111-114.
[100]Chang S,Puryear S,Caimey J.A simple and efficient method for isolating from pine trees.Plant Mol Biol Rep,1993:11:113-116
[101]张维铭.现代分子生物学手册.北京:科学技术出版社,2003
[102]顾红雅,瞿礼嘉,明小天.植物基因与分子操作.北京:北京大学出版社,1995
[103]黄璐琳,胡尚钦,杨晓.药用植物生物工程研究进展Ⅱ.分子标记、遗传转化及功能基因研究.中草药,2006,37(5):647-651
[104]李太武,相建海,刘瑞玉.中国对虾 cDNA 文库的构建.动物学报,1998,44(2):237-238
[105]Sambrook J,Maniatis T,Fristch EF.Molecular cloning:a laboratory manual.(2nd Edition).New York:Cold Spring Harbor Laboratory Press,1989:283-356
[106]王玉成,杨传平,刘桂丰,姜静.紫杆柽柳 cDNA 文库构建与硫氧还蛋白基因的克隆.分子植物育种,2004,2(5):667-673
[107]Audic S and Claverie JM.The significance of digital gene expression profiles.Genome Res,1997,7:986-995
[108]于凤池.EST 技术及其应用综述.中国农学通报,2005,21(2):54-58
[109]崔杰,徐德昌,李滨胜,杨谦,孙璟晗.甜菜 ATP 合酶β亚基基因 atpB 的克隆、序列分析及进化.植物研究,2006,26(5):583-588
[110]Satish CD,Murray LW,Brian KM.Transcriptional activation of cytochrome P450 genes by different classes of chemical inducers.Clin Exp Pharmacol physiol,1998,25:1-9
[111]Kasai H,Nadano D,Hidaka E,Higuchi K,Kawakubo M,Sato TA,Nakayama J.Differential expression of ribosomal protein in human nomal and neop lasticolorectum.J Histochem Cytochem,2003,51:567-574.
[112]熊玲媛,徐金森,陈睦.甜菊钙调蛋白基因的克隆及结构分析.厦门大学学报(自然科学版),2002,41(4):476-480
[113]Helene F,Nicilas C,James D,Pierre-Marie B.Molecular cloning and characterization of calmulin genes in young oak seedlings(Quercus peeraea L.) during early flooding stress.Biochimica et Biophysica Acta,2005,1727:213-219
[114]常团结,朱祯.植物金属硫蛋白研究进展(一).生物技术通报,2002,3:5-10
[115]Mrrs KA.The functions and regulation of glutathione S-transferases in plants.Ann.Rev.Plant Physiol.Plant Mol Biol,1996,47:127-158
[116]Allagulova CR,Gimalov FR,Shakirova FM,Vakh itov VA.The plant dehydrins:structure and putative functions.Biochem,2003,68:945-951.
[117]刘慧丽,李玲.脱落酸(ABA)诱导基因表达的调控元件.植物学通报,2001,18(3):276-282
[118]彭世清,陈守才.植物的遍在蛋白系统.热带农业科学,2001,4:49-54
[119]王金昌,李燕萍.泛素一蛋白酶体途径的生物学功能.江西科学,2006,24(2):155-158
[120]Skinne W,Keon J,Hargreaves J.Gene infoumation for fungal plant pathogens from expressed sequences.Curr Opin Micobiol,2001,4:381-386.
[121]Bevan M,Bancroft I,Bent E,Love K,Goodman H,Dean C,Bergkamp R,Dirkse W,Van Staveren M,StiekemaW.ect.Analysis of 19Mb of contiguous sequences from chromosome 4 of Arabidopsis thaliana.Nature,98,391:53-455
[122]刘关君.EST 技术分析西伯利亚蓼耐盐机制及耐盐相关基因克隆.东北林业大学博士论文,2007
[123]Sanders D,Brownlee C,Haiper JF.Communicating with Calcium.Plant Cell,1999,11,691-706
[124]陈海浪,陈喜文,陈德富.NAD+-依赖型异柠檬酸脱氢酶的结构和功能研究进展.生物技术通报,2003,4(14):304-307
[125]Close T J.Dehydrins:emergence of a biochemical role of a family of plant dehydration proteins.Physiol Plant,1996,97:795-803
[126]Allagulova C R,Gimalov F R,Shakirova F M,Vakh itov VA.The plant dehydrins:structure and putative functions.Biochem,2003,68:945-951.
[127]Nylander M,Svensson J,Palva ET,Welin B V.Stress-in-duced accumulation and tissue- specific localization of dehydrins in A rabid op sis thaliana.Plant Mol Biol,2001,45:263-279
[128]Robertson M,Chandler PM.A dehydrin cognate protein from pea(Pisum sativum L.)with anatypical pattern of ex-pression.Plant Mol Biol,1994,26:805-816.
[129]Stupnikova Ⅳ,Borovsk ii GB,Do rofeev NV,Peshkova AA,Voinikov VK.Accumulation and disappearance of dehydrins and sugars depending on freezing tolerance of winter wheat plants at different developmental phase.J Therm Biol,2002,27:55-60
[130]Liang XW,Dron M,Cramer CL,Dixon RA,Lamb CJ.Differential regulation of phenylalanine ammonia-lyase genes during plant development and by environmental cues.J Biol Chem,1989,264:14486-14492
[131]董艳珍.植物苯丙氨酸解氨酶基因的研究进展.生物技术通报,2006增刊:31-32
[132]Gupta PK,Balyan HS,Sharma PC,Ramesh B.Microsatellites in plants:a new class of molecular markers.Curr Sci,1996,70:45-54
[133]Powell W,Machray GC,Provan J.Polymorphism revealed by simple sequence repeats.Trends Plant Sci,1996,1:215-222
[134]R(o|¨)der MS,Korzun V,Wendehake K,Plashke J,Tixier MH,Leroy P,Ganal MW.A microsatellite map of wheat.Genetics,1998,149:2007-2023
[135]Kantety RV,Rota ML,Matthews DE,Sorrells ME.Data mining for simple sequence repeats in expressed sequence tags from barley,maize,rice,sorghum and wheat).Plant Mol Biol,2002,48:501-510
[136]金基强,李素芳,龚晓春,卢美贞,姚艳玲,忻雅,崔海瑞茶树 EST 资源中 SSR 的信息分析.科技通报,2006,22(4):471-476
[137]李小白,张明龙,崔海瑞.油菜 EST 资源的 SSR 信息分析.中国油料作物学报,2007,29(1):20-25
[138]Cardle L,Ramsay L,Milbourne D,Macaulay M,Marshall D,Waugh R.Computational and experimental characterization of physically clustered simple sequence repeats in plants.Genetics,2000,156:847-854
[139]Morgante M,Hanafey M,Powell W.Microsatellites are prefertially associated with nonrepetitive DNA in plant genomes.Nat genet,2000,30:194-200
[140]李永强,李宏伟,高丽锋,何蓓如.基于表达序列标签的微卫星标记(EST-SSRs)研究进展.植物遗传资源学报,2004,5(1):91-95
[141]Cordeiro GM,Casu R,McIntyre CL,Manners JM,Henry RJ.Microsatellite markers from sugarcane(Saccharum spp.) ESTs cross transferable to erianthus and sorghum.Plant Sci,2001,160:1115-1123
[142]李华盛,范术丽,沈法富.从棉花 ESTs 数据库中筛选微卫星标记的初步研究.棉花学报,2005,17(4):211-216
[143]Eujayl.I,Sledge MK,Wang L,May GD,Chekhovskiy K,Zwonitzer JC,Mian MA.Medicago truncatula EST-SSRs reveal cross-species genetic markers for Medicago spp.Theor Appl Genet,2004,108:414-422
[144]Saha MC,Mian MAR,Eujayl I,Zwonitzer JC,Wang LJ,May GD.Tall fescue EST-SSR markers with transferability across several grass species.Theor Appl Genet,2004,109:783-791
[145]Fraser LG,Harvey CF,Crowhurst RN,DeSilva HN.EST-derived microsatellites from Actinidia species and their potential for mapping.Theor Appl Genet,2004,108:1010-1016
[146]Jung S,Abbott A,Jesudurai C,Tomkins J,Main D.Frequency,type,distribution and annotation of simple sequence reapeats in Rosaceae ESTs.Funct Integr Genomics,2005,5:136-143
[147]Kong Q,Xiang C,Yu Z,Zhang C,Liu F,Peng C and Peng X.Mining and charactering microsatellites in Cucumis melo expressed sequence tags from sequence database.Mol Ecol Notes,2007,7:281-283
[148]忻雅,崔海瑞,卢美贞,姚艳玲,金基强,林容杓,崔水莲.白菜 EST-SSR 信息分析与标记的建立.园艺学报,2006,33(3):549-554
[149]Jia XP,Shi YX,Song YC,Wang GY,Wang TY,Li Y.Development of EST-SSR in foxtail millet(Setaria italica).Genet Resour Crop Evol,2007,54:233-236
[150]陈军方,任正隆,高丽锋,贾继增.从小麦 EST 序列中开发新的 SSR 引物.作物学报,2005,31:154-158
[151]Squirrell J,Hollingsworth P M,Woodhead M,Russell J,Lowe A J,Gibby M and Powell W.How much effort is required to isolate nuclear microsatellites from plants?.Mol Ecol,2003,12:1339-1348
[152]Eujayl I,Sorrells M E,Wolters P and Powell W.Isolation of EST-derived microsatellite markers for genotyping the A and B genomes of wheat.Theor Appl Genet,2002,104:399-407
[153]Scott K D,Eggler P,Seaton G,Rossetto M,Ablett L S and Henry R J.Analysis of SSRs derived from grape ESTs.Theor Appl Genet,2000,100:723-726
[154]Jiang D,Zhong G Y and Hong Q B.Analysis of microsatellites in Citrus unigenes.Acta Genetica Sinica,2006,33:345-353
[2]窦德强,靳玲,陈英杰.人参的化学成分及药理活性的研究进展与展望.沈阳药科大学学报,1999,16(2):151-156
[3]张雅玉.人参皂苷分析方法的研究进展,特产研究,1998,1:50-52
[4]章观德.人参的化学成分和分析方法研究进展,药学学报,1980,15(6):375-384
[5]Ma YC,Luo M,Mally L,Docet M.Distribution and proportion of major ginsenosides and qualty control of ginseng products.Chinese Journal of medical chemistry,1996,6:11-21
[6]柳鹏,李勇.人参皂甙生物活性的研究进展.中国生育健康杂志,2004,15(5):317-319
[7]别光增道.药用人参及其有关植物成分的研究-西洋参的微量皂苷及所谓巴西人参的成分.国外医学中医中药分册,1983,4:42-45
[8]颜春芳,马家锐.人参植株不同部位皂甙含量研究.现代应用药学,1991,8(1):47-48
[9]徐绥绪,陈英杰,吕永俊,王志学,王乃利,姚新生.人参和国产西洋参化学的系统研究.沈阳药学学报,1989,6(4):286-289
[10]郑友兰,张崇禧,李向高.人参和西洋参芦头与根的化学成分比较分析.吉林农业大学人参科技论文集,1988,8:433-437
[11]张翼伸,李润秋,王玉万.人参多糖的研究.东北师范大学学报(Ⅰ),1982,2:97-103
[12]梁忠岩,张翼伸,苗春艳.西洋参与人参总糖组成及含量比较研究.中国药学杂志,1989,24(4):207-209
[13]Ji-Hye Lee and Kyung Hyun Kim.Pectin-like acidic polysaccharides from Panax ginseng with selective antiadhesive activity against pathogenic bacteria.Carbohydrate search,2006,341:1154-1163
[14]沈宁,王素贤,吴立军,李铣.中国吉林栽培西洋参挥发油的新近研究.沈阳药学院学报,1991,8(3):175-181
[151郑友兰,张崇禧,李向高.人参和西洋参芦头与根的化学成分比较分析.吉林农业大学.人参科研论文集,1988.8:433-436
[16]王勇,石磊,王慧,孙允秀,刘在群,罗旭阳,王建辉,林英杰,洪哲.人参茎叶挥发油中倍半萜烯类化合物的分离与鉴定.吉林大学自然科学学报,2001,1:88-90
[17]唐斌,程绪菊,刘江,唐冰.人参皂苷药理作用研究进展.西南军医,2005,7(3):45-47
[18]姜正林,吴新民,陈云,金淑仪,陈燕萍,庄坚.人参皂甙的抗鼠脑缺氧损伤作用.中华航海医学杂志,2000,7(1):28-32
[19]Kim YC,Kim SR,Markelonis GJ,Oh TH.Ginsenosides Rb1and Rg3 protect cultured rat cortical cells from glutamate-induced neurodegeneration.J Neurosci Res,1998,53:426-432
[20]Radad K,Gille G,Moldzio R,Ishige K,Rausch WD.Ginsenosides Rb1 and Rgl effects on survival and neurite growth of MPP(+)-affected mesencephalic dopaminergic cells.J Neural Transm,2004,111:37-45
[21]候明晓,敖定椿.人参总皂甙抗心肌缺血-再灌注损伤的作用机制.中国胸心血管外科临床杂志,2000,7(4):256-259
[22]Okamura N,Kobayashi K,Akaike A,Yagi A.Protective effect of ginseng saponins against impaired brain growth in neonatal rats exposed to ethanol.Biol Pharm Bull,1994,17:270-274
[23]白长喜.人参皂甙 Re 对大鼠心肌缺血-在灌注损伤时心肌和血浆前列腺素 E 水平的影响.延边医学院学报,1993,16(1):17-19
[24]张南生,张秀华,李文峰.人参皂苷 Rg3研究进展.医药导报,2006,25(7):687-689
[25]Mochizuki M,Yoo Yc,Matsuzawa K,Satok K,Saiki I,Tono-oka S,Samukawa K,Azuma I.Inhibitory effect of tumor metastasis in mice by saponins,ginsenoside Rb2,20(R)-and 20(S)-ginsenoside-Rg3,of red ginseng.Biol Pharm Bull,1995,18:197-1202
[26]高勇,王杰军,许青,叶琴琴,郭静,耿怀成.人参皂甙 Rg3抑制肿瘤心神血管形成的研究.第二军医大学学报,2001,22(1):40-42
[27]Nakata H,Kikuchi Y,Tode T,Hirata J,Kita T,Ishii K,Kudoh K,Nagata I,Shinomiya N.Inhibitory effects of ginsenoside Rh2 on tumor growth in nude mice bearing human ovarian cancer cells.Japanese Journal of Cancer Research,1998,89:733-740.
[28]王巍,杜松洁,王乃功.人参皂甙 Rb1和 Rg1对幼小鼠性腺和副性腺的影响.中药药理与临床,1999,15(1):10-11
[29]王艳华,肖雪媛,杨贵贞.人参皂甙对手术应激大鼠下丘脑-垂体-肾上腺轴及免疫靶器官中肾上腺皮质激素释放、ACTH 阳性细胞数的影响.中国神经免疫学和神经病学杂志,1998,5(1):12-15
[30]Haralampidis K,Trojanowska M,Osbourn AE.Biosynthesis of triterpenoid saplonins in plants.Adv Biochem Eng Biotechnol,2002,75:31-49
[31]Walker K,Fuj isaki S,Long R,Croteau R.Molecular cloning and heterologous expression of the C-13 phenylpropanoid side chain-CoA acyltransferase that functions in Taxol biosynthesis.Proc Natl Acad Sci USA,2002,99:12715-12720
[32]Wallaart TE,Bouwmeester HJ,Hille J,Poppinga L,Maijers NC.Amorpha-4,11-diene synthase:Cloning and functional expression of a key enzyme in the biosynthetic pathway of the novel antimalarial drug artemisinin.Planta,2001,212:460-465
[33]Yazaki K,Kunihisa M,Fujisaki T,Sato F.Geranyl diphosphate:4-Hydroxybenzoate geranyltransferase from Lithospermum erythrorhizon.Cloning and characterization of a key enzyme in shikonin biosynthesis.J Biol Chem,2002,277:6240-6246
[34]Grothe T,Lenz R,Kutchan TM.Molecular characterization of the salutaridinol 7-O- acetyltransferase involved in morphine biosynthesis in opium poppy Papaver somniferum.J Biol Chem,2001,276:30717-30723
[35]赵寿经,李昌禹,臧埔,马海琴,杨振堂.影响人参发根中皂甙含量基因的定位及其克隆.特产研究,2001,4:1-5
[36]Bulgakov VP,Khodakovskaya MV,Labetskaya NV,Tchernoded GK,Zhuraclev YN.The impact of plant rolC oncogene on ginsenoside production by ginseng hairy root cultures.Phytochemistry,1998,49:1929-1934
[37]罗志勇,陆秋恒,刘水平,陈湘晖,罗建清,汤立军,胡维新.人参植物皂苷生物合成相关基因的筛选与鉴定.生物化学与生物物理学,2003,35(6):554-560
[38]刘水平,罗志勇,陈湘晖,文斌,汪凌昊,魏来,胡维新.人参皂苷生物合成相关新基因GBR6的 cDNA 克隆及序列分析.生命科学研究,2004,8(4):351-354
[39]王昆,王颖,鲍永利孟祥颖,乌垠,易静文,李玉新.人参叶全长 cDNA 文库的构建及部分克隆的序列分析.分子科学学报,2006,22(1)58-62
[40]胡向阳,张文清,方建颖,蔡伟明,汤章城.脱乙酰壳多糖处理增加人参细胞皂苷的累积和皂苷合成关键酶基因的转录.植物生理与分子生物学学报,2002,28(6):485-490
[41]胡向阳,方建颖,蔡伟明,汤章城.模拟失重对钙调蛋白基因表达与人参皂苷合成的影响.自然科学进展,2005,5(13):537-540
[42]Jung,JD,Park HW,HahnY,Hur CG,In DS,Chung H J,Liu JR,Choi DW.Discovery of genes for ginsenoside biosynthesis by analysis of ginseng expressed sequence tags.Plant Cell Rep,2003,22:224-230
[43]Kim MK,Lee BS,In JG,Sun H,Yoon JH,Yang DK.Comparative analysis of expressed sequence tags(ESTs) of ginseng leaf.Plant Cell Rep,2005,25:599-606
[44]Choi D W,Jung JD,Ha YI,Park HW,In DS,Chung HJ Liu JR.Analysis of transcripts in methyl jasmonate-treated ginseng hairy roots to identify genes involved in the biosynthesis of ginsenosides and other secondary metabolites Plant Cell Rep,2005,23:557-566
[45]Galaud JP,Carri(?)re M,Pauly N,Canut H,Chalon P,Caput D,Pont-ezica RF.Construction of two ordered cDNA libraries enriched in genes encoding plasmalemma and tonoplast proteins from a high-efficiency expression library.Plant J,1999,17:111-118.
[46]Hawkins V,Doll D,Bumgarner R,Smith T,Abajian C,HooD L,Nelson PS.PEDB:the Prostate Expression Database.Nucleic Acids Res,1999,27:204-208
[47]Peterson LA,Brown MR,Carlisle A J,Kohn EC,Liotta LA,Emmert-Buck MR,Krizman DB.An improved method for construction of directionally cloned cDNA libraries from microdissected cells.Cancer Res,1998,58:5326-5328
[48]Diatchenko L,Lau YF,Campbeu AP,Chenchik A,Moqadam F,Huang B,Lukyanov S,Lukyanov K,Gurskaya N,Sverdlov ED,Siebert PD.Suppression subtractive hybridization:a method for generating differentially regulated or tissue-specific cDNA probes and libraries.Proc Natl Acad Sci USA,1996,93:6025-6030
[49]王关林,方宏筠．植物基因工程(第二版)．北京：科学技术出版社,2002：43～170
[50]Young RA and Davis RW.Efficient isolation of genes by using antibody probes.Proc Natl Acad Sci USA,1983,80:1194～1198
[51]Aguan K,Kusano T,Suzuki N,Kitagawa Y.An improved method for the construction of high efficiency cDNA library in plasmid or lambda vector.Nucleic Acids Res,1990,8:1071～1076
[52]Gonos ES,Derventzi A,Kveiborg M,Agiostratidou G,Kassem M,Clar BFC,Jat PS,Rattan SIS.Cloning and identification of genes that associated with mammalian replicative senescence.Exp Cell Res,1998,240:66～74.
[53]Hakvoort TB,Spijkers JA,Vermeulen JL,Lamers WH.Preparation of a differentially expressed full-length cDNA expression library by RecA-mediated triple-strand formation with subtractively enriched cDNA fragments.Nucleic Acids Res,1996,24:3478～3480.
[54]Gurskaya NG,Diatchenko L,Chenchik A,Siebert PD,Khaspekov GL,Lukyanov KA,Vagner LL,Ermolaeva OD,Lukyanov SA,Sverdlov ED.Equalizing cDNA subtraction based on selective suppression of Polymerase Chain Reaction:cloning of Jurkat transcripts induced by phytohemaglutinin and phorbol 12-myristable 13-acetable.Analytical Biochemistry,1996,240:90～97
[55]Wong BR,Rho J,Arron J,Robinson E,Orlinick J,Chao M,Kalachikov S,Cayani E,Bartlett FS 3rd,Frankel WN,Lee SY,Choi Y.Trance is a novel ligand of the tumor necrosis factor receptor family that activates c-Jun N-terminal kinase in T cells.J Biol Chem,1997,272:25190～25194
[56]Chu,ZL,McKinsey TA,Liu L,Gentry JJ,Malim MH,Ballard DW.Suppression of tumour necrosis factor-induced cell death by inhibitor of apoptosis C-IAP2 is under NF-B control.Proc Natl Acad Sci USA,1997,94:10057～10062
[57]Siki RK,Scharf S,Faloona F,Mullis KB,Horn GT,Erlich HA,Arnheim.Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.Science.1985,230:1350～1354
[58]Belyavsky A,Vinogradova T,and Rajewsky K.PCR-based cDNA library construction:general cDNA libraries at the level of a few cells.Nucleic Acids Res,1989,17:2919～2932.
[59]Fromont_Racine M.,Bertrand E,Pictet R,Grange T.A highly sensitive method for mapping the 5' termini of mRNAs.Nucleic Acids Res,1993,21:1683～1684
[60]Suzuki Y,Yoshitomo-Nakagawa K,Maruyama K,Suyama A,Sugano S.Construction and characterization of a full length-enriched and a 5'-end-enriched cDNA library.Gene,1997,200:149～156
[61]Frohman MA,Dush MK,and Martin G.R.Rapid production of full-length cDNAs from rare transcripts:Amplification using a single gene-specific oligonucleotide primer.Proc Natl Acad Sci USA,1988,85:8998-9002
[62]Roeder T.Solid-phase cDNA library construction,a versatile approach.Nucleic Acids Res,1998,26:3451-3452
[63]Rondinelli RH,Tricoli JV.CLARl,a novel gene that exhibits enhanced expression in advanced human prostate cancer.Clin Cancer Res,1999,5:1595-1602
[64]Velculescu VE,Zhang L,Vogelstein B,Kinzler KW.Serial analysis of gene expression.Science,1995,270:484-487
[65]Sompayrac L,Jane S,Bum TC,Tenen DG,and Danna KJ.Overcoming limitations of the mRNA differential display technique.Nucleic Acids Res,1995,23:4738-4739
[66]Adams MD,Kelley JM,Gocayne JD,Dubnick M,Polymeropoulos MH,Xiao H,Merril CR,Wu A,Olde B,Moreno RF.Complementary DNA sequencing:Expressed Sequence Tags and Human Genome Project.Science,1991,252:1651-1656
[67]White JA,Todd J,Newman T,Focks N,Girke T,de Il(?)rduya OM,Jaworski JG,Ohlrogge JB,Benning C.A New set of Arabidopsis Expressed Sequence Tags from Developing Seeds.The etabolic Pathway from Carbohydrates to Seed Oil.Plant Physiol,2000,124:1582-1594
[68]Bouchez D and Hofte H.Functional genomic in plants.Plant Physiology.1998,118:725-732
[69]Ewing RM,Kahla AB,Poirot O.Large-scales statistical analysis-of rice ESTs reveal correlate Kahlad patterns of gene expression.Genome Res,1999,9:950-959
[70]Adams MD,Dubnick M,Kerlavage AR,Moreno R,Kelley JM,Utterback TR,Nagle JW,Fields C,Venter JC.Sequence identification of 2,375 human brain genes.Nature,1992,355:632-634
[71]Asamizu E,Nakamura Y,Sato S,Tabata S.Characteristics of the Lotus japonicus gene repertoire deduced from large-scale expressed sequence tag(EST) analysis.Plant Mol Biol,2004,54:405-414
[72]Wan JS,Sharp S J,Poirier G.M.Cloning differentially expressed mRNA.Nature Biotechnol,1996,14:1685-1691
[73]李靖,韩本立,黄桂君,钱桂生,徐晓霞,杨彤翰,梁平,陈杰.应用抑制消减杂交技术构建肝癌差异表达基因消减 cDNA 文库.第三军医大学学报,2001,23(6):698-701
[74]罗志勇,刘水平,陆秋恒,陈湘晖,文斌,罗建清,胡维新.人参植物与皂苷生物合成相关的差减 cDNA 文库构建及基因差异表达分析.生命科学研究.2003,71(4):324-328
[75]李明芳,郑学勤.开发 SSR 引物方法之研究动态.遗传.2004,26(5):769-776
[76]曾庆国,陈艺燕.微卫星位点筛选方法综述.生态科学,2005,24(4):368-372
[77]Schena M,Schalon D,Davis RW and Brown PQ.Quantitative monito ring of gene expression patterns with a complementary DNA microarray.Science,1995,270:467-470
[78]Schena M.Genome analysis with gene expression microarrays.Bioassays,1996,18:427-431
[79]杨传平,王玉成,刘桂丰,姜静,张国栋.基因芯片技术研究柽柳 NaHCO3 胁迫下基因的表达.生物工程学报,2005,21(2):220-226
[80]祝骥,马文丽,姚汝华.DNA 芯片技术与基因表达研究.生命科学研究.2000,4(2):106-111
[81]Hatey F,Tosser-Klopp G,Clouscard-martinato C,Mulsant P and Gasser F.Expressed sequenced tags for genes:a review.Genet Sel Evol,1998,30:521-541
[82]Zhou Y,Tang JB,Walker MG,Zhang XQ,Wang J,Hu SN.Gene identification and expression analysis of 86136 sequencetags(EST) from the rice genome.Gen Prot Bioinfo,2003,1:26-42
[83]Rossi M,Araujo PG,Paulet F,Garsmeur O,Dias VM,Chen H,Van Sluys MA,D Hont A.Genomic distribution and characterization of EST-derived resistance gene analogs(RGAs)in sugarcane.Mol Gen Genomics,2003,269:406-419
[84]Rounsley S,Linx KK.Large scale sequencing of plant genome.Curr Opin Plant Biol,1998,1:136-141
[85]Sasaki T.The rice genome project in Japan.Proc Natl Acad Sci USA,1998,95:2027-2028
[86]Kurata N,Nagamura Y,Yamamoto K,Sue N,Wu J,Antonio BA,Shomura A,Shimizu T,Lin SY,Inoue T,Fukuda A,Shimano T.Nagamura Y,Yamamoto K,et al.A 300 kilobase interval genetic map of rice including 883 expressed sequences.Nat Genet,1994,8:365-372
[87]Chee PW,Rong J,William-Coplin D.EST derived PCR-based markers for functional gene homologues in cotton.Gene,2004,47:449-462
[88]Kotani H,Nakamura Y,Sato S,Kaneko T,Asamizu E.Structural analysis of Arabidopsis thaliana chromosome 5.VL.Sequence features of the regions of 1,367,185bp covered by 19physically assigned P1 and TAC clones.DNA Res,1998,5:203-216
[89]Schuler GD,Boguski MS,Stewart EA,Stein LD,Gyapay G,Rice K,White RE.ect.A gene map of the human genome.Science,1996,274:540-546
[90]Cui YL,Zhang XY,Zhou Y,Yu H,Tao L,Zhou J,Zhuge Q,Cai YM,Huang MR..Identification and expression Analysis of EST-based Genes in the Bud of Lycoris longituba.Geno Prot Bioinfo,2004,2:42-46
[91]Shin KE,Mary AC,Inna A,Davina B,Monica F,Elizabeth C,John CC.Transcript profiling of salinity stress responses by large-scale expressed sequence tag analysis in Mesembryanthemum crystallinum.Gene,2004,341:83-92
[92]王华春,陈清轩.充分利用 EST 数据库资源.生物化学与生物物理进展,2000,27:442-444
[93]Chen X,Salamini R and Gebhardt C.A Potato molecularfunction map for carbohydrate metabolism and transport.Theor Appl Genet,2001,102:284-295
[94]Thiel T,Miehalek W,Varshney RK,Graner A.Exploiting EST databases for the development and characterization of gen-derived SSR-markers in barley(Hordeum vulgareL.).Theor Appl Genet,2003,106:411-422
[95]Holton RA,Christopher JT,McClure L,Harker N,Henry RJ.Identification and mapping of polymorphic SSR markers from expressed gene sequences of barley and wheat.Mol Breeding,2002,9:63-71
[96]陈亮,赵丽萍,高其康.茶树新梢 cDNA 克隆测序和表达序列标签(ESTs)特性分析.农业生物技术学报,2005,13(1):21-25
[97]王昆,王颖,鲍永利,孟祥颖,乌垠,李玉新.提取高质量人参 RNA 的方法研究.生物化学与生物物理进展,2006,33(1):95-99
[98]王壮伟,渠慎春,章镇,张君毅.苹果属 RNA 高效快速提取新方法.果树学报,2004,21(4):385-387
[99]Wang DH,Wang BC,Li B,Duan CR,Zhang J.Extraction of total RNA from Chrysanthemum containing high levels of phenolic and carbohydrates.Colloid Surface B,2004,36:111-114.
[100]Chang S,Puryear S,Caimey J.A simple and efficient method for isolating from pine trees.Plant Mol Biol Rep,1993:11:113-116
[101]张维铭.现代分子生物学手册.北京:科学技术出版社,2003
[102]顾红雅,瞿礼嘉,明小天.植物基因与分子操作.北京:北京大学出版社,1995
[103]黄璐琳,胡尚钦,杨晓.药用植物生物工程研究进展Ⅱ.分子标记、遗传转化及功能基因研究.中草药,2006,37(5):647-651
[104]李太武,相建海,刘瑞玉.中国对虾 cDNA 文库的构建.动物学报,1998,44(2):237-238
[105]Sambrook J,Maniatis T,Fristch EF.Molecular cloning:a laboratory manual.(2nd Edition).New York:Cold Spring Harbor Laboratory Press,1989:283-356
[106]王玉成,杨传平,刘桂丰,姜静.紫杆柽柳 cDNA 文库构建与硫氧还蛋白基因的克隆.分子植物育种,2004,2(5):667-673
[107]Audic S and Claverie JM.The significance of digital gene expression profiles.Genome Res,1997,7:986-995
[108]于凤池.EST 技术及其应用综述.中国农学通报,2005,21(2):54-58
[109]崔杰,徐德昌,李滨胜,杨谦,孙璟晗.甜菜 ATP 合酶β亚基基因 atpB 的克隆、序列分析及进化.植物研究,2006,26(5):583-588
[110]Satish CD,Murray LW,Brian KM.Transcriptional activation of cytochrome P450 genes by different classes of chemical inducers.Clin Exp Pharmacol physiol,1998,25:1-9
[111]Kasai H,Nadano D,Hidaka E,Higuchi K,Kawakubo M,Sato TA,Nakayama J.Differential expression of ribosomal protein in human nomal and neop lasticolorectum.J Histochem Cytochem,2003,51:567-574.
[112]熊玲媛,徐金森,陈睦.甜菊钙调蛋白基因的克隆及结构分析.厦门大学学报(自然科学版),2002,41(4):476-480
[113]Helene F,Nicilas C,James D,Pierre-Marie B.Molecular cloning and characterization of calmulin genes in young oak seedlings(Quercus peeraea L.) during early flooding stress.Biochimica et Biophysica Acta,2005,1727:213-219
[114]常团结,朱祯.植物金属硫蛋白研究进展(一).生物技术通报,2002,3:5-10
[115]Mrrs KA.The functions and regulation of glutathione S-transferases in plants.Ann.Rev.Plant Physiol.Plant Mol Biol,1996,47:127-158
[116]Allagulova CR,Gimalov FR,Shakirova FM,Vakh itov VA.The plant dehydrins:structure and putative functions.Biochem,2003,68:945-951.
[117]刘慧丽,李玲.脱落酸(ABA)诱导基因表达的调控元件.植物学通报,2001,18(3):276-282
[118]彭世清,陈守才.植物的遍在蛋白系统.热带农业科学,2001,4:49-54
[119]王金昌,李燕萍.泛素一蛋白酶体途径的生物学功能.江西科学,2006,24(2):155-158
[120]Skinne W,Keon J,Hargreaves J.Gene infoumation for fungal plant pathogens from expressed sequences.Curr Opin Micobiol,2001,4:381-386.
[121]Bevan M,Bancroft I,Bent E,Love K,Goodman H,Dean C,Bergkamp R,Dirkse W,Van Staveren M,StiekemaW.ect.Analysis of 19Mb of contiguous sequences from chromosome 4 of Arabidopsis thaliana.Nature,98,391:53-455
[122]刘关君.EST 技术分析西伯利亚蓼耐盐机制及耐盐相关基因克隆.东北林业大学博士论文,2007
[123]Sanders D,Brownlee C,Haiper JF.Communicating with Calcium.Plant Cell,1999,11,691-706
[124]陈海浪,陈喜文,陈德富.NAD+-依赖型异柠檬酸脱氢酶的结构和功能研究进展.生物技术通报,2003,4(14):304-307
[125]Close T J.Dehydrins:emergence of a biochemical role of a family of plant dehydration proteins.Physiol Plant,1996,97:795-803
[126]Allagulova C R,Gimalov F R,Shakirova F M,Vakh itov VA.The plant dehydrins:structure and putative functions.Biochem,2003,68:945-951.
[127]Nylander M,Svensson J,Palva ET,Welin B V.Stress-in-duced accumulation and tissue- specific localization of dehydrins in A rabid op sis thaliana.Plant Mol Biol,2001,45:263-279
[128]Robertson M,Chandler PM.A dehydrin cognate protein from pea(Pisum sativum L.)with anatypical pattern of ex-pression.Plant Mol Biol,1994,26:805-816.
[129]Stupnikova Ⅳ,Borovsk ii GB,Do rofeev NV,Peshkova AA,Voinikov VK.Accumulation and disappearance of dehydrins and sugars depending on freezing tolerance of winter wheat plants at different developmental phase.J Therm Biol,2002,27:55-60
[130]Liang XW,Dron M,Cramer CL,Dixon RA,Lamb CJ.Differential regulation of phenylalanine ammonia-lyase genes during plant development and by environmental cues.J Biol Chem,1989,264:14486-14492
[131]董艳珍.植物苯丙氨酸解氨酶基因的研究进展.生物技术通报,2006增刊:31-32
[132]Gupta PK,Balyan HS,Sharma PC,Ramesh B.Microsatellites in plants:a new class of molecular markers.Curr Sci,1996,70:45-54
[133]Powell W,Machray GC,Provan J.Polymorphism revealed by simple sequence repeats.Trends Plant Sci,1996,1:215-222
[134]R(o|¨)der MS,Korzun V,Wendehake K,Plashke J,Tixier MH,Leroy P,Ganal MW.A microsatellite map of wheat.Genetics,1998,149:2007-2023
[135]Kantety RV,Rota ML,Matthews DE,Sorrells ME.Data mining for simple sequence repeats in expressed sequence tags from barley,maize,rice,sorghum and wheat).Plant Mol Biol,2002,48:501-510
[136]金基强,李素芳,龚晓春,卢美贞,姚艳玲,忻雅,崔海瑞茶树 EST 资源中 SSR 的信息分析.科技通报,2006,22(4):471-476
[137]李小白,张明龙,崔海瑞.油菜 EST 资源的 SSR 信息分析.中国油料作物学报,2007,29(1):20-25
[138]Cardle L,Ramsay L,Milbourne D,Macaulay M,Marshall D,Waugh R.Computational and experimental characterization of physically clustered simple sequence repeats in plants.Genetics,2000,156:847-854
[139]Morgante M,Hanafey M,Powell W.Microsatellites are prefertially associated with nonrepetitive DNA in plant genomes.Nat genet,2000,30:194-200
[140]李永强,李宏伟,高丽锋,何蓓如.基于表达序列标签的微卫星标记(EST-SSRs)研究进展.植物遗传资源学报,2004,5(1):91-95
[141]Cordeiro GM,Casu R,McIntyre CL,Manners JM,Henry RJ.Microsatellite markers from sugarcane(Saccharum spp.) ESTs cross transferable to erianthus and sorghum.Plant Sci,2001,160:1115-1123
[142]李华盛,范术丽,沈法富.从棉花 ESTs 数据库中筛选微卫星标记的初步研究.棉花学报,2005,17(4):211-216
[143]Eujayl.I,Sledge MK,Wang L,May GD,Chekhovskiy K,Zwonitzer JC,Mian MA.Medicago truncatula EST-SSRs reveal cross-species genetic markers for Medicago spp.Theor Appl Genet,2004,108:414-422
[144]Saha MC,Mian MAR,Eujayl I,Zwonitzer JC,Wang LJ,May GD.Tall fescue EST-SSR markers with transferability across several grass species.Theor Appl Genet,2004,109:783-791
[145]Fraser LG,Harvey CF,Crowhurst RN,DeSilva HN.EST-derived microsatellites from Actinidia species and their potential for mapping.Theor Appl Genet,2004,108:1010-1016
[146]Jung S,Abbott A,Jesudurai C,Tomkins J,Main D.Frequency,type,distribution and annotation of simple sequence reapeats in Rosaceae ESTs.Funct Integr Genomics,2005,5:136-143
[147]Kong Q,Xiang C,Yu Z,Zhang C,Liu F,Peng C and Peng X.Mining and charactering microsatellites in Cucumis melo expressed sequence tags from sequence database.Mol Ecol Notes,2007,7:281-283
[148]忻雅,崔海瑞,卢美贞,姚艳玲,金基强,林容杓,崔水莲.白菜 EST-SSR 信息分析与标记的建立.园艺学报,2006,33(3):549-554
[149]Jia XP,Shi YX,Song YC,Wang GY,Wang TY,Li Y.Development of EST-SSR in foxtail millet(Setaria italica).Genet Resour Crop Evol,2007,54:233-236
[150]陈军方,任正隆,高丽锋,贾继增.从小麦 EST 序列中开发新的 SSR 引物.作物学报,2005,31:154-158
[151]Squirrell J,Hollingsworth P M,Woodhead M,Russell J,Lowe A J,Gibby M and Powell W.How much effort is required to isolate nuclear microsatellites from plants?.Mol Ecol,2003,12:1339-1348
[152]Eujayl I,Sorrells M E,Wolters P and Powell W.Isolation of EST-derived microsatellite markers for genotyping the A and B genomes of wheat.Theor Appl Genet,2002,104:399-407
[153]Scott K D,Eggler P,Seaton G,Rossetto M,Ablett L S and Henry R J.Analysis of SSRs derived from grape ESTs.Theor Appl Genet,2000,100:723-726
[154]Jiang D,Zhong G Y and Hong Q B.Analysis of microsatellites in Citrus unigenes.Acta Genetica Sinica,2006,33:345-353
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