Abstract:A strain of Mycobacterium tuberculosis H37Rv with Hsp16.3 gene knock-out were constructed in the study. Mycobacterium tuberculosis strain H37Rv was cultured in vitro and the genome DNA was extracted. In order to obtain a gene targeting vector pKO-Hsp16.3, two fragments (5′-Hsp16.3 and 3′-Hsp16.3) flanking the Hsp16.3 region were amplified by PCR and inserted into pKO plasmid, and the recombinant pKO-Hsp16.3 plasmid was obtained. Gene exchange took place within the genome of Mycobacterium tuberculosis H37Rv after pKO-Hsp16.3 plasmid was transformed into it, and the strain of Mycobacterium tuberculosis H37Rv with Hsp16.3 gene knock-out was selected. Results indicated that the target strain was the positive one identified by two step polymerase chain reaction (PCR) and one step sucrose counter selection simultaneously and could not grow in hygomycin culture media. Finally, the strain of Mycobacterium tuberculosis H37Rv with Hsp16.3 gene knock-out is constructed successfully in our laboratory.
田玺择,吴 芳,章 乐,曹旭东,张万江. 结核分枝杆菌H37Rv菌株Hsp16.3基因缺失突变株的构建与鉴定[J]. 中国人兽共患病学报, 2013, 29(10): 991-995.
TIAN Xi-ze,WU Fang,ZHANG Le,CAO Xu-dong,ZHANG Wan-jiang. Construction and identification on a strain of Mycobacterium tuberculosis H37Rv with Hsp16.3 gene knock-out. Chinese Journal of Zoonoses, 2013, 29(10): 991-995.
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