脓肿分枝杆菌非溶血性磷脂酶C蛋白MAB_0555基因克隆测序及生物信息学分析

目的克隆脓肿分枝杆菌（Mycobacterium abscessus,M.abscessus）非溶血性磷脂酶C基因MAB_0555,并对其测序以及生物信息学分析,为进一步研究其生物学功能及其在脓肿分枝杆菌致病机制中的作用提供基础。方法以脓肿分枝杆菌基因组DNA为模板,PCR扩增非溶血性磷脂酶C的基因片段,克隆入pQE-30质粒,构建重组原核表达载体pQE-30-MAB_0555,将其转化入大肠埃希菌DH5α,并进行序列测定及利用生物信息学软件DNAstar 5.0分析其生物学特性。结果 PCR扩增获得长1440 bp的MAB_0555基因片段,编码479个氨基酸,DNAstar等软件预测其蛋白质相对分子量（Mr）约为53 kD,并显示出具有良好的抗原性。结论成功获得序列正确的脓肿分枝杆菌MAB-0555基因,为其重组蛋白的表达及其相关研究奠定基础。

Cloning,sequencing and analysis of the biological information of non-hemolytic PLC MAB0555 gene of Mycobacterium abscessus

Objective To clone and sequence non-hemolytic PLC MAB-0555 gene of M.abscessus,analyze its biological function,so as to provide scientific basis for further research on its biological function and pathogenic mechanism.MethodThe MAB-0555 gene was amplified from the DNA of M.abscessus by PCR and cloned into vector pQE-30.The constructed recombinant plasmid pQE-30-MAB₀555 was transformed to competent E.coli DH5α and sequenced.The DNAstar 5.0 was used to analyze its biological properties at the amino acids level.Result The MAB₀555 gene of 1 440 bp,coding the polypeptides of 479 amino acids,was amplified by PCR.The biological information software predicted its relative molecular mass（Mr） was about 53 kD and possessed good antigencity.Conclusion A comfirmed MAB₀555 gene has been cloned,which provided a good foundation for its recombination,expression and related study.