中国现代医学杂志2006，Vol.16Issue(10)：1441-1444,4.
人前列腺特异性抗原基因重组表达载体pPICZαC-m PSA的构建和表达
Construction and expression of recombinant human prostate-specific antigen expression vector pPICZα-mPSA
邹冬辉 1张家颖 2郭焱 1路英丽 1左文静 1王忠山1
作者信息
- 1. 吉林大学基础医学院细胞生物学教研室,吉林,长春,130021
- 2. 吉林大学基础医学院生物化学与分子生物学实验中心,吉林,长春,130021
- 折叠
摘要
Abstract
[Objective] To produce biologically active recombinant human (rh) prostate-specific antigen in a Pichia pastoris for clinic detection and therapeutic use. [Methods] The cDNA encoding the full length human PSA was obtained by RT-PCR and was sequenced. By inserting cDNA encoding mature hPSA 237 amino acid residues into Pichia pastoris expression vector pPICZα-C consisting of AOX1 promoter and the yeast α-factor signal sequence, the recombinant expression plasmid pPICZα-mPSA was constructed and transformed to yeast host strain X33. The protein was induced to express by methanol and the one transformant which expressed rPSA in high level was screened out by ELISA Kit. [Results] The cDNA encoding the full human PSA was obtained and the expression plasmid pPICZα-mPSA was constructed and the X33 cell strain which expressed rPSA in high level was obtained, and the final yield of active rPSA from Pichia pastoris was 1.2mg/L. [Conclusion] A method of expression of mature PSA in Pichia pastoris has been established. Large-scale production of active rPSA will be useful in the study of PSA biology character, clinic detection and therapeutic use.关键词
前列腺特异性抗原/毕赤酵母Key words
prostate-specific antigen/pichia pastoris分类
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邹冬辉,张家颖,郭焱,路英丽,左文静,王忠山..人前列腺特异性抗原基因重组表达载体pPICZαC-m PSA的构建和表达[J].中国现代医学杂志,2006,16(10):1441-1444,4.
