单增李斯特氏菌溶血素基因的克隆及原核表达
Cloning and Prokaryotic Expression of Hemolysin Gene of Listeria monocytogenes
  
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中文摘要:
      
英文摘要:
      In this study, the Hly gene ofListeria monocytogenes train C53005 was amplified by PCR using designed primers. Then, the gene was cloned into pMD18-T vector, and indentified by digestion with restriction endonuclease, PCR and sequencing. After the sequences were confirmed correct, a prokaryotic expression vector was constructed by inserting the gene into pET-28a vector. The recombinant plasmid pET-28a- silly was transformed into E.coli BL21 (DE3) competent cells, and was induced with 1PTG. The expressed product was analysed by SDS-PAGE and Western-blot. In result, the Hly gene was highly expressed in E.coli and the expressed protein was 65 Ku in size as expected. Western-blot test demonstrated that the expressed protein could specifically react with rabbit anti-LM serum. The results showed the expressed protein was in soluble form with satisfactory antigenicity.
作者单位
吴晓薇,徐成刚,马保华,江经纬,叶贺佳,区燕宜,徐小芹,廖明  
中文关键词:  单核细胞增生性李斯特氏菌  溶血素基因  克隆  原核表达
英文关键词:Listeria monocytogenes  Hemolysin  Gene cloning  Prokaryotic expression
基金项目:广东出入境检验检验局科技项目
DOI:
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