摘要
目的 将副溶血性弧菌直接溶血毒素 6 5位氨基酸进行突变。方法 酶切 tdh基因获得tdhm2 6 8- 5 70 bp,一对突变引物扩增出 tdhm 1- 2 6 7bp,与载体 p U C19连接并转化大肠杆菌 JM10 9;对重组质粒进行测序鉴定。结果 DNA测序结果表明 tdhm和 tdh的序列有 2个碱基的差别。结论 按照预期设计成功突变了副溶血性弧菌的直接溶血毒素的 6 5位氨基酸 。
Objective To mutate the 65 th amino acid(AA) of the thermostable direct hemolysin(TDH) of Vibrio parahaemolyticus(Vp). Methods Digestion with the restriction endonucleases BamHI and HincⅡ was performed to get the fragments tdhm268 570 bp. PCR technique with a couple of mutation primers was adopted to amplify the fragments tdhm1 267 bp which were digested by the restriction endonucleases EcoRI and HincⅡ. The digested fragments were ligated with plasmid pUC19. The recombinant plasmid was subsequently transformed into E.coli JM109, and was identified and sequenced. Results There were only two bp difference between the sequence of the positive clone and the reported tdh. Conclusions The 65 th AA of TDH had been successfully mutated,which was paid the preliminary foundation to construct the vaccine of Vp.
出处
《疾病控制杂志》
2002年第4期301-302,共2页
Chinese Journal of Disease Control and Prevention
