结核分枝杆菌H37 Ra菌株ESAT6基因的克隆及序列分析

摘要

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.%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。