结核分枝杆菌H37Ra菌株ESAT6基因的克隆及序列分析

Cloning and sequential analysis of ESAT6 gene of Mycobacterium tuberculosis H37Ra

摘要: 目的: 从结核分枝杆菌H37Ra菌株克隆早期分泌性抗原靶6(ESAT6)基因并进行序列分析.方法: 以结核分枝杆菌H37Ra菌株基因组DNA为模版,用PCR方法扩增ESAT6基因,将扩增产物连接到克隆载体pTG19-T中,并用PCR、单双酶切和测序进行鉴定,以BLAST软件进行序列比对分析.结果: 从结核分枝杆菌H37Ra株成功克隆了ESAT6基因,测序表明该片段由288 bp组成,与GenBank所报告的H37Rv基因相比同源性为100%.结论: 成功克隆了H37Ra株ESAT6编码基因,其序列与H37Rv标准株ESAT6编码基因完全一致.

Abstract: Objective: To clone and sequence the ESAT6 gene from Mycobacterium tuberculosis(MTB) H37Ra.Methods: The encoding gene of ESAT6 was amplified from MTB H37Ra genomic DNA using PCR technique.The amplified PCR product was subcloned into pTG19-T vector.The target gene was identified by PCR,single and double enzyme digestion,sequencing and sequence alignment analysis with BLAST software.Results: The ESAT6 gene was successfully cloned.The sequencing showed the fragment length was 288 bp,which was homology with the MTB H37Ra gene reported by GenBank.Conclusions: The coding gene of ESAT6 from MTB H37Ra is successfully cloned.