PNP基因克隆及其表达载体的构建与鉴定
摘要
目的:研究PNP基因的克隆、真核表达载体的构建与鉴定,为肿瘤的基因治疗与基因工程的应用提供实验和理论依据。
方法:①大肠埃希氏杆菌K12 PNP基因的PCR扩增;②重组质粒pMSCV-PNP的构建;③用氯化钙法制备感受态大肠杆菌(XL1-Blue);④转化感受态菌:用携带目的基因的重组质粒pMSCV-PNP转化感受态XL1-Blue大肠杆菌,同时设立阳性对照组和阴性对照组。转移适量转化菌于平皿培养,筛选氨苄青霉素抗性转化菌落;⑤提取质粒和酶切重组质粒:提取重组质粒pMSCV-PNP,限制性内切酶EcoRⅠ和BglⅡ双酶切重组质粒,电泳鉴定;⑥PCR鉴定、测序鉴定重组质粒pMSCV-PNP;⑦提取质粒和酶切质粒:限制性内切酶BspHⅠ、BglⅡ、AflⅡ分别酶切pVSV-G,pGAG-POL,pMSCV,电泳鉴定质粒。
结果:①PNP基因PCR扩增出特异条带;②重组质粒pMSCV-PNP电泳结果与预期一致;③重组质粒pMSCV-PNP转化结果:实验组平皿上可见近百个菌落,阳性对照组有数百个菌落生长,而阴性对照组则未见菌落生长;④阳性克隆质粒pMSCV-PNP PCR鉴定、酶切鉴定,电泳条带与预期一致;测序结果正常;⑤克隆质粒pVSV-G,pGAG-POL,pMSCV酶切鉴定与预期一致。
结论:①从大肠埃希氏杆菌K12中克隆到PNP基因;②构建了表达载体pMSCV-PNP;③抽提了三种载体质粒pMSCV、pVSV-G、pGAG-POL。
Objective: To construct and identify the recombinant vector pMSCV-PNP carrying E.coli K12 PNP which can express in eukaryote cells and which will provide the basis for gene therapy.
Methods:①PNP were amplified from E.coli K12 bacteria by polymerase chain reaction (PCR).②To construct the recombinant vector pMSCV-PNP by means of T4-DNA ligase.③Competent E. coli (strain XL1-Blue) was prepared using calcium chloroid.④Transformation of the competent bacteria: We transformed the competent bacteria with the plasmids pVSV-G, pGAG-POL, pMSCV and pMSCV-PNP containing PNP gene. The appropriate volume of the transformed bacteria was transfered and spreaded onto agar LB medium to select the ampicillin-resistant colonies.⑤Extraction and restriction endonuclease analysis of the plasmids: The plasmids pVSV-G, pGAG-POL, pMSCV, pMSCV-PNP were extracted from the transformed bacteria with plasmid purification kit, and they were cut with restriction endonucleases BspHⅠ、BglⅡ、AflⅡ, EcoRⅠand BglП, respectively.
Results:①Ampicillin resistant tranformed bacteria colonies growth: After transformation with pMSCV , pMSCV-PNP, pGAG-POL and pVSV-G, about 100 colonies were observed on the plate containing ampicillin, and the colonies were not found in the negative control groups.②The normal size of the fragments from the plasmids cut by endonucleases: The results of restriction endonuclease analysis and agar electrophoresis indicated that size of plasmids pMSCV, pMSCV-PNP, pGAG-POL and pVSV-G was normal.
Conclusion: E.coli K12 PNP can be successfully cloned and inserted into expression vector. The newly constructed vector may serve as the potential tool to conduct further comprehensive experiments in future on PNP function and on gene therapy.
方法:①大肠埃希氏杆菌K12 PNP基因的PCR扩增;②重组质粒pMSCV-PNP的构建;③用氯化钙法制备感受态大肠杆菌(XL1-Blue);④转化感受态菌:用携带目的基因的重组质粒pMSCV-PNP转化感受态XL1-Blue大肠杆菌,同时设立阳性对照组和阴性对照组。转移适量转化菌于平皿培养,筛选氨苄青霉素抗性转化菌落;⑤提取质粒和酶切重组质粒:提取重组质粒pMSCV-PNP,限制性内切酶EcoRⅠ和BglⅡ双酶切重组质粒,电泳鉴定;⑥PCR鉴定、测序鉴定重组质粒pMSCV-PNP;⑦提取质粒和酶切质粒:限制性内切酶BspHⅠ、BglⅡ、AflⅡ分别酶切pVSV-G,pGAG-POL,pMSCV,电泳鉴定质粒。
结果:①PNP基因PCR扩增出特异条带;②重组质粒pMSCV-PNP电泳结果与预期一致;③重组质粒pMSCV-PNP转化结果:实验组平皿上可见近百个菌落,阳性对照组有数百个菌落生长,而阴性对照组则未见菌落生长;④阳性克隆质粒pMSCV-PNP PCR鉴定、酶切鉴定,电泳条带与预期一致;测序结果正常;⑤克隆质粒pVSV-G,pGAG-POL,pMSCV酶切鉴定与预期一致。
结论:①从大肠埃希氏杆菌K12中克隆到PNP基因;②构建了表达载体pMSCV-PNP;③抽提了三种载体质粒pMSCV、pVSV-G、pGAG-POL。
Objective: To construct and identify the recombinant vector pMSCV-PNP carrying E.coli K12 PNP which can express in eukaryote cells and which will provide the basis for gene therapy.
Methods:①PNP were amplified from E.coli K12 bacteria by polymerase chain reaction (PCR).②To construct the recombinant vector pMSCV-PNP by means of T4-DNA ligase.③Competent E. coli (strain XL1-Blue) was prepared using calcium chloroid.④Transformation of the competent bacteria: We transformed the competent bacteria with the plasmids pVSV-G, pGAG-POL, pMSCV and pMSCV-PNP containing PNP gene. The appropriate volume of the transformed bacteria was transfered and spreaded onto agar LB medium to select the ampicillin-resistant colonies.⑤Extraction and restriction endonuclease analysis of the plasmids: The plasmids pVSV-G, pGAG-POL, pMSCV, pMSCV-PNP were extracted from the transformed bacteria with plasmid purification kit, and they were cut with restriction endonucleases BspHⅠ、BglⅡ、AflⅡ, EcoRⅠand BglП, respectively.
Results:①Ampicillin resistant tranformed bacteria colonies growth: After transformation with pMSCV , pMSCV-PNP, pGAG-POL and pVSV-G, about 100 colonies were observed on the plate containing ampicillin, and the colonies were not found in the negative control groups.②The normal size of the fragments from the plasmids cut by endonucleases: The results of restriction endonuclease analysis and agar electrophoresis indicated that size of plasmids pMSCV, pMSCV-PNP, pGAG-POL and pVSV-G was normal.
Conclusion: E.coli K12 PNP can be successfully cloned and inserted into expression vector. The newly constructed vector may serve as the potential tool to conduct further comprehensive experiments in future on PNP function and on gene therapy.
引文
[1] Beerens AM, Rots MG, Bermúdez B, et al. Secretion of thymidine kinase to increase the effectivity of suicide gene therapy results in the loss of enzymatic activity[J]. Journal of Drug Targeting, 2008, 16(1):26-35.
[2] Bertin S, Neves S, Gavelli A, et al. Cellular and molecular events associated with the antitumor response induced by the cytosine deaminase/5-fluorocytosine suicide gene therapy system in a rat liver metastasis model[J]. Cancer Gene Therapy, 2007, 14 (10):858-866.
[3] Pugmire MJ, Ealick SE. Structural analyses reveal two distinct families of nucleoside phosphorylases[J]. The Biochemical Journal, 2002, 361 (Pt 1):1-25.
[4] Bzowska A, Kulikowska E, Shugar D. Purine nucleoside phosphorylases: properties, functions, and clinical aspects[J]. Pharmacology & Therapeutics, 2000, 88 (3):349-425.
[5] Modrak-Wójcik A, Stepniak K, Akoev V, et al. Molecular architecture of E. coli purine nucleoside phosphorylase studied by analytical ultracentrifugation and CD spectroscopy[J]. The Protein Society, 2006, 15(7):1794-1800.
[6] Parker WB, King SA, Allan PW. In vivo gene therapy of cancer with E.coli purine nucleoside phosphorylase[J]. Human gene therapy, 1997, 8 (14): 1637-1644.
[7] Hong JS, Waud WR, Levasseur DN, et al. Excellent in vivo bystander activity of fludarabine phosphate against human glioma xenografts that express the Escherichia coli purine nucleoside phosphorylase gene[J]. Cancer Research, 2004, 64 (18):6610-6615.
[8] Burns JC, Friedmann T, Driever W, et al. Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vector: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells[J]. Proc Natl Acad Sci U S A, 1993, 90 (17): 8033-8037.
[9] 张兆山. 附录2培养基[M]. 见:黄培堂主译. 分子克隆实验指南,第三版.北京:科学出版社,2002, 1595-1598.
[10] 张兆山. 附录2培养基[M]. 见:黄培堂主译. 分子克隆实验指南,第三版.北京:科学出版社,2002, 1598-1599.
[11] Williams SR, Goddard JM, Martin DW Jr. Human purine nucleoside phosphorylase cDNA sequence and genomic clone characterization[J]. Nucleic acids research, 1984, 12(14): 5779–5787.
[12] 张兆山. 附录2培养基[M]. 见:黄培堂主译. 分子克隆实验指南,第三版. 北京:科学出版社,2002, 1599-1600.
[13] 吕星,瞿祥虎. 质粒及其在分子克隆中的应用[M]. 见:黄培堂主译.分子克隆实验指南,第三版. 北京:科学出版社,2002, 96-98.
[14] 吕星,瞿祥虎. 质粒及其在分子克隆中的应用[M]. 见:黄培堂主译.分子克隆实验指南,第三版. 北京:科学出版社,2002, 91.
[15] 吕星,瞿祥虎. 质粒及其在分子克隆中的应用[M]. 见:黄培堂主译.分子克隆实验指南,第三版. 北京:科学出版社,2002, 98-99.
[16] Daly G, Chernajovsky Y. Recent developments in retroviral-mediated gene transduction[J]. Molecular Therapy, 2000, 2(5): 423-434.
[17] Anson DS. Retroviral-mediated gene transduction[J]. Methods in molecular biology,2001, 175: 471-494.
[18] Persons DA, Allay JA, Allay ER, et al. Retroviral-mediated transfer of the green fluorescent protein gene into murine hematopoietic cells facilitates scoring and selection of transduced progenitors in vitro and identification of genetically modified cells in vivo[J]. Blood, 1997, 90(5):1777-1786.
[19] De Miguel MP, Donovan PJ. Determinants of retroviral-mediated gene delivery to mouse spermatogonia[J]. Biology of reproduction, 2003, 68(3):860-866.
[20] Barrette S, Douglas J, Orlic D, et al. Superior transduction of mouse hematopoietic stem cells with 10A1 and VSV-G pseudotyped retrovirus vectors[J]. Molecular Therapy, 2000, 1(4): 330-338.
[21] 石小玉,李文林,梁朝,等. 重组小鼠干细胞逆转录病毒载体高效基因转染CD41+、UT7、U937和MDA-MB-435细胞[J].中国病理生理杂志,2004, 20(2):212-218.
[22] 唐镇生主编. 分子外科与基因治疗[M]. 第一版. 上海:上海医科大学出版社,1999,9-25.
[23] Zhu XL, Kumar R, Mandal M, et al. Cell cycle-dependent modulation of telomerase activity in tumor cells[J]. Proc Natl Acad Sci U S A.1996, 93 (12):6091 -6095.
[1] Beerens AM, Rots MG, Bermúdez B, et al. Secretion of thymidine kinase to increase the effectivity of suicide gene therapy results in the loss of enzymatic activity[J]. Journal of Drug Targeting, 2008,16(1):26-35.
[2] Bertin S, Neves S, Gavelli A, et al. Cellular and molecular events associated with the antitumor response induced by the cytosine deaminase/5-fluorocytosine suicide gene therapy system in a rat liver metastasis model[J]. Cancer Gene Therapy, 2007,14(10):858-866.
[3] Pugmire MJ, Ealick SE. Structural analyses reveal two distinct families of nucleoside phosphorylases[J]. The Biochemical Journal, 2002,361(Pt 1):1-25.
[4] Bzowska A, Kulikowska E, Shugar D. Purine nucleoside phosphorylases: properties, functions, and clinical aspects[J]. Pharmacology & Therapeutics, 2000,88(3):349-425.
[5] NCBI. deoD purine-nucleoside phosphorylase [ Escherichia coli K12 ] [OL]. 10/1/2008. http://www.ncbi.nlm.nih.gov/sites/entrez?Db=gene&Cmd=ShowDetailView&TermToSearch=945654&ordinalpos=7&itool=EntrezSystem2.PEntrez.Gene.Gene_ResultsPanel.Gene_RVDocSum.
[6] NCBI. NP nucleoside phosphorylase [ Homo sapiens ] [OL]. 10/1/2008. http://www.ncbi.nlm.nih.gov/sites/entrez?Db=gene&Cmd=ShowDetailView&TermToSearch=4860&ordinalpos=17&itool=EntrezSystem2.PEntrez.Gene.Gene_ResultsPanel.Gene_RVDocSum..
[7] Parker WB, Allan PW, Shaddix SC, et al. Metabolism and metabolic actions of 6-methylpurine and 2-fluoroadenine in human cells[J]. Biochemical Pharmacology, 1998,55(10):1673-1681.
[8] Mackey JR, Baldwin SA, Young JD, et al. Nucleoside transport and its significance foranticancer drug resistance[J]. Drug Resistance Updates, 1998;1(5):310-324.
[9] Hong JS, Waud WR, Levasseur DN, et al. Excellent in vivo bystander activity of fludarabine phosphate against human glioma xenografts that express the Escherichia coli purine nucleoside phosphorylase gene[J]. Cancer Research, 2004,64(18):6610-6615.
[10] Sorscher EJ, Peng S, Bebok Z, et al. Tumor cell bystander killing in colonic carcinoma utilizing the Escherichia coli DeoD gene to generate toxic purines[J]. Gene Therapy, 1994, 1(4):233-238.
[11] Rubsam LZ,Boucher PD ,Murphy PJ, et al. Cytotoxicity and accumulation of ganciclovir triphosphate in bystander cells cocultured with herpes simplex virus type 1 thymidine kinase-expressing human glioblastoma cells[J]. Cancer Research, 1999,59(3):669-675.
[12] Li Z ,Shanmu GN ,Kata YD, et al. Enzyme/ prodrug gene therapy approach for breast cancer using a recombinant adenovirus expressing Escherichia coli cytosine deaminase[J]. Cancer Gene Therapy, 1997,4(2):113-117.
[13] Deharvengt S, Wack S, Uhring M, et al. Suicide gene/prodrug therapy for pancreatic adenocarcinoma by E. coli purine nucleoside phosphorylase and 6-methylpurine 2′-deoxyriboside[J]. Pancreas, 2004,28(2):E54-64.
[14] Zhou JH, Tang B, Liu XL, et al. hTERT-targeted E. coli purine nucleoside phosphorylase gene/6-methylpurine deoxyribose therapy for pancreatic cancer[J]. Chinese Medical Journal, 2007,120(15): 1348-1352.
[15] 黄颖,曾岩,郑永晨.大肠杆菌嘌呤核苷磷酸化酶自杀基因对人胃癌细胞的作用[J].吉林大学学报(医学版), 2007,33(1):105-108.
[16] Kikuchi E, Menendez S, Ozu C, et al. Delivery of replication-competent retrovirus expressing Escherichia coli purine nucleoside phosphorylase increases the metabolism of the prodrug, fludarabine phosphate and suppresses the growth of bladder tumor xenografts[J]. Cancer Gene Therapy, 2007 ,14(3):279-286.
[17] Martiniello-Wilks R, Wang XY, Voeks DJ, et al. Purine nucleoside phosphorylase and fludarabine phosphate gene-directed enzyme prodrug therapy suppresses primary tumour growth and pseudo-metastases in a mouse model of prostate cancer[J]. The Journal of Gene Medicine, 2004,6(12):1343-1357.
[18] Ungerechts G, Springfeld C, Frenzke ME,et al. Lymphoma chemovirotherapy: CD20-targeted and convertase-armed measles virus can synergize with fludarabine[J]. Cancer Research, 2007,67(22):10939-10947.
[19] Bennett EM, Anand R, Allan PW, et al. Designer gene therapy using an Escherichia coli purine nucleoside phosphorylase/prodrug system[J]. Chemistry & Biology, 2003,10(12): 1173-1181.
[20] Yang Z, William BP, Eric J, et al. PNP Anticancer Gene Therapy[J]. Current Topics in Medicinal Chemistry, 2005,5(13):1259-1274.
[2] Bertin S, Neves S, Gavelli A, et al. Cellular and molecular events associated with the antitumor response induced by the cytosine deaminase/5-fluorocytosine suicide gene therapy system in a rat liver metastasis model[J]. Cancer Gene Therapy, 2007, 14 (10):858-866.
[3] Pugmire MJ, Ealick SE. Structural analyses reveal two distinct families of nucleoside phosphorylases[J]. The Biochemical Journal, 2002, 361 (Pt 1):1-25.
[4] Bzowska A, Kulikowska E, Shugar D. Purine nucleoside phosphorylases: properties, functions, and clinical aspects[J]. Pharmacology & Therapeutics, 2000, 88 (3):349-425.
[5] Modrak-Wójcik A, Stepniak K, Akoev V, et al. Molecular architecture of E. coli purine nucleoside phosphorylase studied by analytical ultracentrifugation and CD spectroscopy[J]. The Protein Society, 2006, 15(7):1794-1800.
[6] Parker WB, King SA, Allan PW. In vivo gene therapy of cancer with E.coli purine nucleoside phosphorylase[J]. Human gene therapy, 1997, 8 (14): 1637-1644.
[7] Hong JS, Waud WR, Levasseur DN, et al. Excellent in vivo bystander activity of fludarabine phosphate against human glioma xenografts that express the Escherichia coli purine nucleoside phosphorylase gene[J]. Cancer Research, 2004, 64 (18):6610-6615.
[8] Burns JC, Friedmann T, Driever W, et al. Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vector: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells[J]. Proc Natl Acad Sci U S A, 1993, 90 (17): 8033-8037.
[9] 张兆山. 附录2培养基[M]. 见:黄培堂主译. 分子克隆实验指南,第三版.北京:科学出版社,2002, 1595-1598.
[10] 张兆山. 附录2培养基[M]. 见:黄培堂主译. 分子克隆实验指南,第三版.北京:科学出版社,2002, 1598-1599.
[11] Williams SR, Goddard JM, Martin DW Jr. Human purine nucleoside phosphorylase cDNA sequence and genomic clone characterization[J]. Nucleic acids research, 1984, 12(14): 5779–5787.
[12] 张兆山. 附录2培养基[M]. 见:黄培堂主译. 分子克隆实验指南,第三版. 北京:科学出版社,2002, 1599-1600.
[13] 吕星,瞿祥虎. 质粒及其在分子克隆中的应用[M]. 见:黄培堂主译.分子克隆实验指南,第三版. 北京:科学出版社,2002, 96-98.
[14] 吕星,瞿祥虎. 质粒及其在分子克隆中的应用[M]. 见:黄培堂主译.分子克隆实验指南,第三版. 北京:科学出版社,2002, 91.
[15] 吕星,瞿祥虎. 质粒及其在分子克隆中的应用[M]. 见:黄培堂主译.分子克隆实验指南,第三版. 北京:科学出版社,2002, 98-99.
[16] Daly G, Chernajovsky Y. Recent developments in retroviral-mediated gene transduction[J]. Molecular Therapy, 2000, 2(5): 423-434.
[17] Anson DS. Retroviral-mediated gene transduction[J]. Methods in molecular biology,2001, 175: 471-494.
[18] Persons DA, Allay JA, Allay ER, et al. Retroviral-mediated transfer of the green fluorescent protein gene into murine hematopoietic cells facilitates scoring and selection of transduced progenitors in vitro and identification of genetically modified cells in vivo[J]. Blood, 1997, 90(5):1777-1786.
[19] De Miguel MP, Donovan PJ. Determinants of retroviral-mediated gene delivery to mouse spermatogonia[J]. Biology of reproduction, 2003, 68(3):860-866.
[20] Barrette S, Douglas J, Orlic D, et al. Superior transduction of mouse hematopoietic stem cells with 10A1 and VSV-G pseudotyped retrovirus vectors[J]. Molecular Therapy, 2000, 1(4): 330-338.
[21] 石小玉,李文林,梁朝,等. 重组小鼠干细胞逆转录病毒载体高效基因转染CD41+、UT7、U937和MDA-MB-435细胞[J].中国病理生理杂志,2004, 20(2):212-218.
[22] 唐镇生主编. 分子外科与基因治疗[M]. 第一版. 上海:上海医科大学出版社,1999,9-25.
[23] Zhu XL, Kumar R, Mandal M, et al. Cell cycle-dependent modulation of telomerase activity in tumor cells[J]. Proc Natl Acad Sci U S A.1996, 93 (12):6091 -6095.
[1] Beerens AM, Rots MG, Bermúdez B, et al. Secretion of thymidine kinase to increase the effectivity of suicide gene therapy results in the loss of enzymatic activity[J]. Journal of Drug Targeting, 2008,16(1):26-35.
[2] Bertin S, Neves S, Gavelli A, et al. Cellular and molecular events associated with the antitumor response induced by the cytosine deaminase/5-fluorocytosine suicide gene therapy system in a rat liver metastasis model[J]. Cancer Gene Therapy, 2007,14(10):858-866.
[3] Pugmire MJ, Ealick SE. Structural analyses reveal two distinct families of nucleoside phosphorylases[J]. The Biochemical Journal, 2002,361(Pt 1):1-25.
[4] Bzowska A, Kulikowska E, Shugar D. Purine nucleoside phosphorylases: properties, functions, and clinical aspects[J]. Pharmacology & Therapeutics, 2000,88(3):349-425.
[5] NCBI. deoD purine-nucleoside phosphorylase [ Escherichia coli K12 ] [OL]. 10/1/2008. http://www.ncbi.nlm.nih.gov/sites/entrez?Db=gene&Cmd=ShowDetailView&TermToSearch=945654&ordinalpos=7&itool=EntrezSystem2.PEntrez.Gene.Gene_ResultsPanel.Gene_RVDocSum.
[6] NCBI. NP nucleoside phosphorylase [ Homo sapiens ] [OL]. 10/1/2008. http://www.ncbi.nlm.nih.gov/sites/entrez?Db=gene&Cmd=ShowDetailView&TermToSearch=4860&ordinalpos=17&itool=EntrezSystem2.PEntrez.Gene.Gene_ResultsPanel.Gene_RVDocSum..
[7] Parker WB, Allan PW, Shaddix SC, et al. Metabolism and metabolic actions of 6-methylpurine and 2-fluoroadenine in human cells[J]. Biochemical Pharmacology, 1998,55(10):1673-1681.
[8] Mackey JR, Baldwin SA, Young JD, et al. Nucleoside transport and its significance foranticancer drug resistance[J]. Drug Resistance Updates, 1998;1(5):310-324.
[9] Hong JS, Waud WR, Levasseur DN, et al. Excellent in vivo bystander activity of fludarabine phosphate against human glioma xenografts that express the Escherichia coli purine nucleoside phosphorylase gene[J]. Cancer Research, 2004,64(18):6610-6615.
[10] Sorscher EJ, Peng S, Bebok Z, et al. Tumor cell bystander killing in colonic carcinoma utilizing the Escherichia coli DeoD gene to generate toxic purines[J]. Gene Therapy, 1994, 1(4):233-238.
[11] Rubsam LZ,Boucher PD ,Murphy PJ, et al. Cytotoxicity and accumulation of ganciclovir triphosphate in bystander cells cocultured with herpes simplex virus type 1 thymidine kinase-expressing human glioblastoma cells[J]. Cancer Research, 1999,59(3):669-675.
[12] Li Z ,Shanmu GN ,Kata YD, et al. Enzyme/ prodrug gene therapy approach for breast cancer using a recombinant adenovirus expressing Escherichia coli cytosine deaminase[J]. Cancer Gene Therapy, 1997,4(2):113-117.
[13] Deharvengt S, Wack S, Uhring M, et al. Suicide gene/prodrug therapy for pancreatic adenocarcinoma by E. coli purine nucleoside phosphorylase and 6-methylpurine 2′-deoxyriboside[J]. Pancreas, 2004,28(2):E54-64.
[14] Zhou JH, Tang B, Liu XL, et al. hTERT-targeted E. coli purine nucleoside phosphorylase gene/6-methylpurine deoxyribose therapy for pancreatic cancer[J]. Chinese Medical Journal, 2007,120(15): 1348-1352.
[15] 黄颖,曾岩,郑永晨.大肠杆菌嘌呤核苷磷酸化酶自杀基因对人胃癌细胞的作用[J].吉林大学学报(医学版), 2007,33(1):105-108.
[16] Kikuchi E, Menendez S, Ozu C, et al. Delivery of replication-competent retrovirus expressing Escherichia coli purine nucleoside phosphorylase increases the metabolism of the prodrug, fludarabine phosphate and suppresses the growth of bladder tumor xenografts[J]. Cancer Gene Therapy, 2007 ,14(3):279-286.
[17] Martiniello-Wilks R, Wang XY, Voeks DJ, et al. Purine nucleoside phosphorylase and fludarabine phosphate gene-directed enzyme prodrug therapy suppresses primary tumour growth and pseudo-metastases in a mouse model of prostate cancer[J]. The Journal of Gene Medicine, 2004,6(12):1343-1357.
[18] Ungerechts G, Springfeld C, Frenzke ME,et al. Lymphoma chemovirotherapy: CD20-targeted and convertase-armed measles virus can synergize with fludarabine[J]. Cancer Research, 2007,67(22):10939-10947.
[19] Bennett EM, Anand R, Allan PW, et al. Designer gene therapy using an Escherichia coli purine nucleoside phosphorylase/prodrug system[J]. Chemistry & Biology, 2003,10(12): 1173-1181.
[20] Yang Z, William BP, Eric J, et al. PNP Anticancer Gene Therapy[J]. Current Topics in Medicinal Chemistry, 2005,5(13):1259-1274.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |