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Position: Home > Articles > Clone and expression of thermostable direct hemolysin of Vibrio parahaemolyticus Journal of Shanghai Ocean University 2009,18 (3) 257-262

副溶血弧菌耐热直接溶血毒素基因的克隆及原核表达

作  者:
丁莹;潘迎捷;赵勇;孙晓红;金维荣;徐晓晶;秦红友
单  位:
上海生物芯片有限公司;上海海洋大学食品学院
关键词:
副溶血弧菌;耐热直接溶血毒素;克隆;纯化蛋白
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。
译  名:
Clone and expression of thermostable direct hemolysin of Vibrio parahaemolyticus
作  者:
DING Ying1,PAN Ying-jie1,ZHAO Yong1,SUN Xiao-hong1,JIN Wei-rong2,XU Xiao-jing2,QIN Hong-you2(1.College of Food Science and Technology,Shanghai Ocean University,Shanghai 201306,China;2.Shanghai Biochip Co.Ltd,Shanghai 201203,China)
关键词:
Vibrio parahaemolyticus;thermostable direct hemolysin;clone;purified protein
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 protein had hemolytic activity.

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