文章摘要
丁莹,潘迎捷,赵勇,孙晓红,金维荣,徐晓晶,秦红友.副溶血弧菌耐热直接溶血毒素基因的克隆及原核表达[J].上海海洋大学学报,2009,(3):.
DING Ying,PAN Ying-jie,ZHAO Yong,SUN Xiao-hong,JIN Wei-rong,XU Xiao-jing,QIN Hong-you.Clone and expression of thermostable direct hemolysin of Vibrio parahaemolyticus[J].Journal of Shanghai Ocean University,2009,(3):.
副溶血弧菌耐热直接溶血毒素基因的克隆及原核表达
Clone and expression of thermostable direct hemolysin of Vibrio parahaemolyticus
  
DOI:
中文关键词: 副溶血弧菌  耐热直接溶血毒素  克隆  纯化蛋白
英文关键词: Vibrio parahaemolyticus  thermostable direct hemolysin  clone  purified protein  
基金项目:上海市科技兴农重点攻关项目
作者单位
丁莹,潘迎捷,赵勇,孙晓红,金维荣,徐晓晶,秦红友  
摘要点击次数: 2009
全文下载次数: 108
中文摘要:
      根据GenBank上已有的副溶血弧菌耐热直接溶血毒素的基因序列,设计并人工合成引物.将耐热直接溶血毒素的基因全长克隆到大肠杆菌表达载体pGEX- 4T-1上,构建重组表达载体tdhA/pGEX- 4T-1和tdhS/pGEX- 4T-1,转化大肠杆菌BL21(DE3),得到表达的工程菌株.优化诱导表达条件,表达耐热直接溶血毒素.转化有重组质粒tdhA/pGEX- 4T-1,tdhS/pGEX- 4T-1的BL21可稳定高效地表达可溶形式的目的蛋白.表达产物用聚丙烯酰胺凝胶电泳鉴定,用GST琼脂糖珠柱亲和层析纯化.溶血实验表明,表达的蛋白具有溶血活性.
英文摘要:
      According to the gene sequence of thermostable direct hemolysin in GenBank,thermostable direct hemolysin gene was subcloned into an Escheria coli expression vector pGEX-4T-1.Recombined plasmids tdhA/pGEX-4T-1,tdhS/pGEX-4T-1 were then transformed into Escheria coli BL21(DE3).The recombinant plasmid tdhA/pGEX-4T-1 and tdhS/pGEX-4T-1 had stable and efficient expression in BL21,the high pure fused protein could be obtained by using affinity chromatography method.The result of hemolytic test showed that the prot...
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