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重组截短型人源结缔组织生长因子在大肠杆菌中的表达

Expression of truncated recombinant human connective tissue growth factor in E.coli
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摘要 用RT-PCR法从人胎盘组织中克隆出人源结缔组织生长因子(h-CTGF)cDNA序列867bp,将此cDNA亚克隆至表达载体pET-9a,重组质粒转化BL21(DE3)pLysS,诱导表达出N端缺失61个氨基酸残基的截短型rh-CTGF,表达量占总菌体蛋白的7%,主要以不溶性包涵体形式存在。采用离心、洗涤和凝胶过滤分离纯化后,行活性检测表明截短型rh-CTGF无刺激增殖活性,并对用原核表达rh-CTGF作了讨论.为制备抗体、CTGF表达调控和功能研究打下了基础。 Connective tissue growth factor(CTGF)emerges in both physiological and pathological processes and may lead to fibrosis,which provides a new target for therapeutic intervention.A867bp cDNA fragment of human-CTGF was amplified from human placenta by RT-PCR and subcloned into the expression vector pET9a.Then it was trans-formed into E.coli strain BL21(DE3)pLsS.The truncated recombinant h-CTGF lacked N terminal61aa and mainly occured in inclusion bodys.rh-CTGF was isolated and purified and could not stimulate the proliferation of fibroblast.The strategy of CTGF expression in prokaryotic cells was discussed.All above will be useful for antibody preparation against CTGF and further research of CTGF expression regulation and anti-fibrosis intervention.
出处 《生物技术通讯》 CAS 2002年第3期164-166,共3页 Letters in Biotechnology
关键词 人结缔组织生长因子 RT-PCR 重组表达 大肠杆菌 截短型rh-CTGF human connective tissue growth factor RT-PCR recombination and expression E.coli
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  • 1Gradlham DM,Igarshi A,Potter RL,et al.Connective tissue growth factor:a cysteine-rich mitogen secreted by human vascular endothelial cells is related to the SRC-induced immediate early gene product CEF-10[J].J Cell Biol,1991,114(6):1285.
  • 2Oemar BS,Luscher JF.Connective tissue growth factor:Friend or foe[J].Art Eriosder Thromb Vasc Biol,1997,17(8):1483.
  • 3Perbal B.NOV(nephroblastoma overexpressed)and the CCN growth factor is genes:structural and functional issues[J].Mol Pathol,2001,54(2):57
  • 4Oemar BS,Werner A,Garnier JM,Human connective tissue growth factor is expressed in advanced atherosclerotic lesions[J].Circulation,1997,95(4):831
  • 5Sambrook J,Fritsch EF,Maniatis T.Molecular Cloning-A Laboratory Manual.2nd ed [M].New York:Cold Spring Harbor,1989
  • 6Bollag DM,Rozycki MD,Edelstein SJ.Protein Method.2nd ed[M].New York:Wiley-Liss Inc,1996
  • 7李琦涵,赵红玲,和绍清,王丽春,赵树栋,王炯.人结缔组织生长因子(CTGF)的原核表达研究[J].中国生物化学与分子生物学报,1999,15(5):833-835. 被引量:3
  • 8Kim HS,Nagalla SR,Oh Y,et al.Identification of a family of low-affinity insulin-like growth factor binding proteins(IGF-BPs):characterization of connective tissue growth factor as a member of the IGFBP superfamily[J].Proc Natl Acad Sci USA,1997,94(24):12981
  • 9Fan WH,Pech M,Karnovsky MJ.Connective tissue growth factor(CTGF)stimulates vascular smooth muscle cell growth and migration in vitro[J].Eur J Cell Biol,2000,79(12):915
  • 10Igarashi A,Okochi H,Bradham H,et al.Regulation of connective tissue growth factor gene expression in human skin fibroblasts and during wound repair[J].Mol Biol Cell,1993,4:637

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