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两个新型多拷贝毕赤酵母表达载体的构建 被引量：4

Construction of two new multi-copy Pichia expression vectors
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摘要 目的 :构建合适的载体用于外源基因在巴斯德毕赤酵母中的正确分泌表达。方法 :利用PCR技术 ,对已有载体pPIC9K进行改造 :在多克隆位点处增加一个NaeⅠ酶切位点 ;去除卡那霉素抗性基因中的一个XhoⅠ酶切位点。结果和结论 :成功地构建了pMEN9K ,pMEX9K两个载体 。 Objective:To construct suitable vectors for the secretory expression of heterologous proteins in Pichia pastoris .Methods:A new unique restriction site Nae Ⅰ was added in multiple cloning sites and a restriction site Xho Ⅰ was eliminated in kanamycin resistance gene.Results and Conclusions:Two new vectors pMEN9K and pMEX9K derived from pPIC9K were successfully constructed,and they are suitable for the secretory expression of heterologous proteins.
作者 齐连权 陈薇 于长明 来大志 翁少洁 王海涛
机构地区 军事医学科学院微生物流行病研究所
出处 《军事医学科学院院刊》 CSCD 北大核心 2002年第4期264-266,共3页 Bulletin of the Academy of Military Medical Sciences
关键词 巴斯德毕赤酵母 载体 基因表达 PCR Pichia pastoris expression vector
分类号 Q782 [生物学—分子生物学]
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参考文献4

  • 1[1]Cregg JM, Cereghino JL, Shi JY, et al. Recombinant protein expression in Pichia pastoris[J]. Mol Biotechnol, 2000,16(1):23-52.
  • 2[2]Goda S,Takano K, Yamagata Y, et al. Effect of extra N-terminal residues on the stability and folding of human lysozyme expressed in Pichia pastoris[J].Protein Eng, 2000, 13(4):299-307.
  • 3[3]Singh A,Lugovoy JM,Kohr WJ, et al. Synthesis, secretion and processing of α-factor-interferon fusion proteins in yeast[J].Nucleic Acids Res,1984, 12 (23):8927-8938.
  • 4[4]Cook AL,Kirwin PM, Craig S, et al. Purification and analysis of proteinase-resistant mutants of recombinant platelet-derived growth factor-BB exhibiting improved biological activity[J]. Biochem J,1992,281(Pt 1):57-65.

同被引文献40

  • 1叶秀珍,金健健,徐链,慕晓军,李文裕.外源血小板衍生生长因子(PDGF)B链基因在CHO细胞的表达[J].实验生物学报,1989,22(3):313-323. 被引量：2
  • 2Hauptmann R Swetly P.A novel class of human type Ⅰ interferons[J].Nucleic Acids Res,1985,13(13):4739-4749.
  • 3Blatt L M, Davis J M, Klein S B, Taylor M W. The biologic activity and molecular characterization of a novel synthetic intederon-alpha species, consensus intederon. J Interferon Cytokine Res, 1996, 16(7) :489 ~ 499.
  • 4Ozes O N, Reiter Z, Klein S, Blatt L M, Taylor M W. A comparison of intederon-Con1 with natural recombinant intederons-alpha: antiviral,antiproliferative, and natural killer-inducing activities. J Interferon Res.1992, 12( 1 ) :55 ~ 59.
  • 5Koyama A H, Arakawa T, Adachi A. Comparison d an antiviral activity of recombinant consensus interferon with recombinant intedcron-alpha-2b. Microbes Infect, 1999, 1(13): 1073 ~ 1077.
  • 6Klein M L, Bartley T D, Davis J M, Whiteley D W, Lu H S. Isolation and Structural characterization of three isoforms of recombinant consensus alpha interferon. Arch Biochem Biophys. 1990, 276(2) : 531 ~ 537.
  • 7Cregg J M, Vedvick T S, Raschke W C. Recent advances in the expression of foreign genes in Pichia pastoris. Biotechnology (N Y),1993,11(8) :905 ~ 910.
  • 8Lin Cereghino G P, Sunga A J, Lin Cereghino J, Cregg J M.Expression of foreign genes in the yeast Pichia pastoris. Genet Eng ( NY). 2001, 23:157 ~ 169.
  • 9Cregg J M, Cereghino J L, Shi J, Higgins D R Recombinant protein expression in Pichia pastoris. Mol Biotechnol, 2000, 16( 1 ) : 23 ~ 52.
  • 10Klein S B, Blatt L M, Taylor M W. Cell surface binding characteristics correlate with consensus type I interferon enhanced activity. J Interferon Cytokine Res. 1996,16(1) : 1 ~ 6.

引证文献4

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军事医学科学院院刊
2002年 第4期

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