分枝杆菌噬菌体CJAUS9 lysB基因的克隆与序列分析

Cloning and Sequence Analysis of Mycobacteriophage CJAUS9 lysB

参照GenBank中lysB基因序列设计特异引物,以耻垢分枝杆菌噬菌体CJAUS9的基因组为模板,PCR扩增出1029 bp的条带,经酶切、PCR鉴定含有目的基因的克隆载体pMD19-T正确后进行测序分析。结果显示,克隆出的lysB基因与已知分枝杆菌肌尾噬菌体lysB序列同源性为98.7%～99.1%;编码的氨基酸同源性为99%～100%,有酯酶保守的G-X-S-X-G基序,为研究LysB蛋白的生物学特性奠定了基础。

Specific primers were designed according to sequences of myoviridae mycobacteriophage mycolylarabinogalactan esterase gene （lysB） in GenBank. Target bands were amplified by PCR with Mycobacterium smegmatis phage CJAUS9 genome as a template, connected with the cloning vector pMD19 - T and transformed into E. coli DHSct. Recombinant plasmids were extracted, digested, identificated by PCR and sequenced. Results show 1029bp lysB gene was successfully cloned and 98.7% to 99.1% sequence homology with other myoviridae mycobacteriophages＇ s lysB and 99% to 100% amino acid homology. G - X - S - X - G esterase conservative motif was also found. The research laid the foundation for bionomics of LysB.