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凝胶色谱柱复性在低分子量尿激酶原突变体(DscuPA-32K)中的应用 被引量:4

Application of a Gel Chromatography Renaturing Way on Low Molecular Singlechain Urokinase Mutant(DscuPA-32K)
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摘要 将构建的一种具溶栓和抗栓双重功能尿激酶原突变体 (DscuPA 32K)基因 ,在大肠杆菌中进行表达。由于DscuPA 32K分子较大并且表达量较高 ,目的蛋白质基本以包涵体的形式存在。包涵体中的蛋白质是无活性的蛋白质 ,为了获得有活性的蛋白质 ,就需要对包涵体进行变性及复性。尝试了一种新的凝胶色谱柱复性方法 ,并通过柱复性方法与常规的稀释复性方法进行了比较 ,发现柱复性方法明显优于稀释复性方法 ,具有成本低 ,效率高 ,并对目的蛋白质 (DscuPA 32K)进行了初步纯化等优点 ,尤其对酶这一类容易失活降解的蛋白质进行复性时 。 A recombinant mutant gene with thrombolytic and antithrombolytic bifunction was expressed in \%E.coli\%. Owing to two reasons of high molecular weight and over expression, dscuPA existed in inclusion body form. The protein of inclusion body was inactive protein. In order to obtain active protein, inclusion bodies should be denatured and then renatured. We performed a novel way named gel\|chromatography column renaturation way. Compare with traditional renaturation way, this refolding approach had some obvious advantages, such as low cost and high recovery, and accomplished the preliminary purification step of desired protein(DscuPA\|32K). Especially to proteins that easily became inactive and degradation, this approach might have good prospect.
出处 《生物工程学报》 CAS CSCD 北大核心 2001年第3期300-303,共4页 Chinese Journal of Biotechnology
基金 国家高技术研究发展计划项目! ( 86 3 10 2 0 9)&&
关键词 低分子量尿激酶原 包涵体 复性 基因突变体 色谱柱洗脱 溶栓药物 low molecular single chain urokinase, inclusion bodies, renaturation
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  • 1沈雳,陈少萍,蔡在龙,杨生生,秦永文.嵌合水蛭肽的构建与活性分析[J].中国生物化学与分子生物学报,2004,20(5):702-706. 被引量:4
  • 2宁保安,马茹,郑玉玲,高志贤,申博,姜永强.RGD-葡激酶突变体(K130T,K135R)的制备与活性分析[J].生物工程学报,2005,21(3):456-460. 被引量:5
  • 3高基民,郑仲承,林来新妹,王小宁,刘新垣.白细胞介素—2—绿脓杆菌外毒素融合蛋白的分高纯化及其复性研究[J].生物化学与生物物理学报,1996,28(1):70-76. 被引量:6
  • 4Panda,A K,Bioprocessing of the therapeutic proteins from the inclusion bodies of Escherichia coli[J].Adv Biochem Eng Biotechnol,2003,85:43~93
  • 5Hevehan D L,Clark E D B.Oxidative renaturation of lysozyme at high concentration[J].Biotechnol Bioeng.
  • 6Xie Y,Wetlaufer D B.Control of aggregation in protein folding:the temperature-leap tactic[J].Protein Sci,1996,5:517~523.
  • 7Walsh,G,Biopharmaceutical benchmarks[J].Nat Biotechnol 2003,21:865~870
  • 8Isumoto,K,Ejima,D,Kumagai,I,Practical considerations in refolding proteins from inclusion bodies[J].Protein Expr Purif.2003,28:1~8
  • 9Derman A I,Prinz W,Belin D et al.Mutations that allow disulfide bond formation in the cytosol of Escherichia coli[J].Sciene,1993,262:1744~1747
  • 10Lili H,Schwarz E,Rudolph R.Advances in refolding of proteins produced in E.coli[J].Curr Opin Biotechnol,1998,9:497~501

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