结核分枝杆菌的aceA基因的克隆、序列测定及生物信息学分析

Cloning and bioinformatic analysis of aceA from M.tb H37Rv

目的 克隆编码结核分枝杆菌异柠檬酸裂合酶的基因aceA (1915 ,1916 ) ,对其全序列进行测定,并以生物信息学方法和软件分析其生物学特性,为进一步研究该酶的功能提供信息资料。方法 用PCR系统扩增出异柠檬酸裂合酶基因片段,将其插入载体pET2 8a,对其全序列进行测定;利用序列对比分析、蛋白氨基酸序列分析及结构预测软件进行生物信息学分析和模拟。结果 克隆的基因序列与报道的序列完全一致,aceA与icl及其他编码异柠檬酸裂合酶基因同源性很高,具有保守氨基酸共有序列;初步模拟出了该蛋白的三级结构。结论 本研究通过对aceA基因序列和氨基酸进行生物信息学分析,获取了该酶的信息特征,并与现有的报道结果进行对比分析,为分子生物学诊断、治疗和药靶筛选提供理论依据。

Objective To acquire and analyze aceA gene from M. tuberculosis H37Rv strain genomic DNA and so as to search it's role as new drug target or diagnose candidate for preventing, controlling and treating Tuberculosis.Methods In this experiment, the aceA gene was amplified by polymerase chain reaction (PCR). Then its genome sequence and encoding proteins was analyzed by BLAST and DNA TOOL,ClustalX,nnPredict,PredictProtein software.Results The sequence of aceA cloning was same as that of published by Genbank. Bioinformatics analysis proved that the aceA gene shares highly conserved sequence and nucleotides with icl gene of M.tb.Conclusion By carry out sequencing and bioinformatics of it analysis,our research established foundation of further explore for Isocitrate lyase enzyme as a potential drug target.