Acta Scientiarum Naturalium Universitatis Pekinensis

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Construction and Expression of the Gene for Human Proinsulin Analog (B-Arg-Arg-A)

LI Xiongbiao, XIE Bin, CHEN Laitong, TANG Jianguo, HU Meihao   

  1. College of Life Sciences, Peking University, Beijing, 100871
  • Received:1992-11-16 Online:1995-01-20 Published:1995-01-20



  1. 北京大学生命科学学院, 北京,100871

Abstract: By means of deletion mutagenesis by polymerase chain reactio (PCR), a plasmid, pUC19BR2A, was constructed which contained the codons of ananalog(B-Arg-Arg-A) of human proinsulin in which the normal35-amino acid connecting peptide was replaced by dipeptide(Arg-Arg). An expression plasmid with PRPL promoter, pJG107, was constructedwhich encoded a fusion protein consisting of B-Arg-Arg-A and a polypeptide of 72 amino acid. Expressed in Eschericbia coli, the fusion protein with insulin immunoreactive activity measured by radioimmunoassay accounted for 58% of total bacterial protein.

Key words: proinsulin analog, gene expression, deletion mutagenesis, polymerase chain reaction

摘要: 用聚合酶链反应(PCR)法将C肽为6个氨基酸残基的胰岛素原类似物基因删除突变成C肽仅为2个精氨酸残基的胰岛素原类似物(BArg-Arg-A)基因,即把pUC18BC'A改建为pUC18BR2A。再将pUC18BR2A与pJG105重组为表达质粒pJG107,使B-Arg-Arg-A与pJG105编码的一种多肽构成融合蛋白在大肠杆菌体系中表达。融合蛋白占细菌蛋白总量的58%。表达产物具有人胰岛素放射免疫活性。

关键词: 胰岛素原类似物, 聚合酶链反应, 基因表达, 删除突变

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