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黑曲霉GA Ⅰ基因的克隆及在P.pastoris中的分泌表达 被引量：2

Secretory Expression of GAI gene in The Pichia pastoris
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摘要 采用RT-PCR技术,从黑曲霉的菌丝体RNA中扩增出葡萄糖淀粉酶GAI的结构基因,将其连接到pPICZαA载体中,转入巴斯德毕赤酵母GS115中,获得8株重组酵母;甲醇诱导目的蛋白分泌表达,葡萄糖淀粉酶酶活最高达174U/mL,占上清液蛋白的36%。 Amplified gene of Glucoamylase Ⅰ from total RNA of Aspergillus niger mycelium by RT-PCR technology, inserted it into pPICZαA vector.Then we transformed the recombinant plasim into Pichia pastoris GSll5 by electroporation, and got 8 recombinant strains. The activity of Glucoamylase Ⅰ was maximum 174U/mL in BMMY medium by methanol induction. Glucoamylase Ⅰ account for 36% of total protein in media supernate. Effect of multiple copies and prefer codon on efficiency of secretory expression was also discussied in detail.
作者 郭德军 柳增善
机构地区 黑龙江八一农垦大学食品学院 吉林大学畜牧兽医学院
出处 《黑龙江八一农垦大学学报》 2005年第4期69-72,共4页 journal of heilongjiang bayi agricultural university
关键词 黑曲霉 葡萄糖淀粉酶 毕赤酵母 分泌表达 Aspergillus niger Glucoamylase Pichia pastoris secretory expression
分类号 Q781 [生物学—分子生物学]
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参考文献13

  • 1Sauer J, Sigurskjold BW, Christensen U, et al. Glucoamylase: structure/function relationships, and protein engineering[J]. Biochim Biophys Acta, 2000,29(2):275-293.
  • 2Cregg JM, Cereghino JL, Shi J, et al. Recombinant protein expression in Pichia pastoris[J]. Mol Biotechnol, 2000,16(1):23-52.
  • 3Boel E, Hjort I, Svensson B, et al. Glucoamylases G1 and G2 from Aspergillus niger are synthesized from two different but closely related mRNAs[J].EMBO J.,1984,3(5):1097-102.
  • 4张树政 孟广震 何忠效.酶学研究技术[M].北京：科学出版社,1987..
  • 5Fierobe HP,Mirgorodskaya E, Frandsen TP, et al. Overexpression and characterization of Aspergillus awamori wild-type and mutant glucoamylase secreted by the methylotrophic yeast Pichia pastoris:comparison with wild-type recombinant glucoamylase produced using Saccharomyces cerevisiae and Aspergillus niger as hosts[JJ.Protein Expr Purif.,1997,9(2):159-170.
  • 6Cregg JM,Barringer KJ, Hessler hY, et al. Pichia pastoris as a host system for transformations[J].Mol Cell Biol, 1995,(5):3376-3385.
  • 7Sreekrish K, Nelles L, Potenz RH, et al. High-level expression, purification, and characterization of recom-binant human tumor factor synthesized in the methylotropic yeast Pichia pastoris[J]. Biochemistry, 1989,(28):4117-4125.
  • 8Clare JJ, Romanos MA, Henwood CA. Production of mouse epidermal growth factor in yeast:high-level secretion using pichia pastoris strains containing multiple gene copies[J].Gene, 1991,(105):205-212.
  • 9姚斌,张春义,王建华,范云六.高效表达具有生物学活性的植酸酶的毕赤酵母[J].中国科学（C辑）,1998,28(3):237-243. 被引量：79
  • 10Thill GP, Davis GR, Stillman GR, et al. Positiv and negative affects of multi-copy integrated expressionvectors on protein expression in Pichia pastoris[A].Proceeding of the sixth international symposium on genetic of industrial microorganisms[C].Pairs: Societe Franscaise de Microbiologie, 1990,447-490.

共引文献97

同被引文献25

  • 1金华利,张富春,单文娟,张爱莲,李轶杰,王宾.口蹄疫病毒VP1蛋白在酵母中的表达及免疫原性分析[J].细胞与分子免疫学杂志,2004,20(5):513-516. 被引量：10
  • 2骆诗鸿.巴斯德毕赤酵母高效表达外源蛋白的策略[J].海峡药学,2007,19(2):4-7. 被引量：3
  • 3刘兆玺,杜金华,王晓晓,马明.安卡红曲霉液态发酵产色素及糖化酶的研究[J].食品与发酵工业,2007,33(4):77-81. 被引量：15
  • 4Valle'e H.,Carre' H.. a pluralite du virus aphteux C.R. Hebd [ J ].Acad.Sci.Paris, 1922,174: 1498-1500.
  • 5Su C X, Duan X G, Wang X Q, et al. Heterologous expression of FMDV immunodominant epitopes and HSP70 in P. pastoris and the subsequent immune response in mice [J ]. Veterinary Microbiology, 2007,124 : 256-263.
  • 6Schneider J,Gilbert S C,Blanchard T J,et al.Enhanced immunogenicity for CD8 +Tcell induction and complete protective efficacy of malaria DNA vaccination by boosting with modified vaccinia virus Ankara [J].Nat Med, 1998,4 (4) : 397-408.
  • 7Kokuho T, Watanabe S, Yokamizo Y,et al.Production of biologically active ,heterodimer-ic porcine interleukin-12 using a monocistronicbaculoviral expression system [J]. Vet Immunol Immunopathol, 1999,72(34):289-302.
  • 8Holmgren J, Lycke N, Czerkinsky C. Cholera toxin and Cholera toxin B subunit as oral-mucosal adjuvant and antigen vector systems [ J ].Vaccine, 1993,11 ( 12 ) : 1184- 1197.
  • 9Yamamoto M, Mcghee J R, Hagiwara Y H, et al.Genetically manipulated bacterial toxin as a new generation mucosal adjuvant[J]. Scand J Immunol, 2001, 53(3): 211-217.
  • 10Faber KN, Hader W, Ab G,et al. Methylotrophic yeasts as factories for the production of foreign proteins[J ].Yeast, 1995, 11(14):1331-1334.

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黑龙江八一农垦大学学报
2005年 第4期

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