摘要
目的:用基因工程方法克隆产毒性大肠杆菌(ETEC)不耐热肠毒素(LT)和耐热肠毒素(ST)基因,以制备检测ETEC的单克隆抗体。方法:利用PCR技术扩增LT和ST基因并将它们插入T-easy载体中,采用全自动测序仪进行核苷酸序列分析。结果:DNA序列分析表明,克隆的ETEC-LT-B DNA序列与GenBank公布序列比较在17、69、101、102位点有碱基变异,同源性98·9%;ETEC-STDNA序列与GenBank公布序列一致。结论:成功获得正确的ETEC的LT和ST基因,为其重组表达及后续研究奠定了实验基础。
Objective ： Heat-stable enterotoxin （ST） and heat-labile enterotoxin （LT） genes in enterotoxigenic Escherichia coli （ETEC） were cloned with gene engineering technique and automatic sequencer was used to analyze their sequences for purpose of producing a monoclonal antibody against ETEC in the future. Methods：ST and LT genes were amplified with PCR and then cloned into T-easy vector. Automatic sequencer was used to analyze their sequences. Results：The sequencing results showed that compared with the GenBank,four mutant base pair were found in the sequence of ETEC-LT-B, being localizd at 17,69,101,102 sites,respectively. The sequence of ETEC-ST was consistent with that in GenBank. Conclusions：The true sequences of ST and LT genes were obtained in the present study and this laid a strong experimental foundation for the further research of His-ST and His-LT proteins expressions.
出处
《华北国防医药》
2005年第5期305-306,共2页
Medical Journal of Beijing Military Region
基金
首都医学发展科研基金项目
项目编号2003-3-55
