结核分枝杆菌Rv0354c基因的克隆及生物信息学分析  

Cloning and bioinformatics analysis of Rv0354c from Mycobacterium tuberculosis

作　　者：李敏英 王楚彤 袁美丽 李柏青[1,2] 许涛 汪洪涛[1,2] LI Minying;WANG Chutong;YUAN Meili;LI Baiqing;XU Tao;WANG Hongtao(Anhui Province Key Laboratory of Immunology in Chronic Diseases.Bengbu Medical College,Bengbu 233030,China;Department of Immunology,School of Laboratory Medicine,Bengbu Medical College,Bengbu 233030,China;Department of Clinical Laboratory,School of Laboratory Medicine,Bengbu Medical College,Bengbu 233030,China)

机构地区：[1]蚌埠医学院慢性疾病免疫学基础与临床安徽省重点实验室,蚌埠233030 [2]蚌埠医学院免疫学教研室,蚌埠233030 [3]蚌埠医学院临床检验诊断学教研室,蚌埠233030

出　　处：《生命的化学》2021年第12期2686-2694,共9页Chemistry of Life

基　　金：安徽省自然科学基金项目(1908085MH252,2008085QH405)。

摘　　要：从NCBI中获得PPE7蛋白的编码基因序列,设计引物从H37Rv基因组中克隆出Rv0354c基因,将其转化到大肠杆菌感受态细胞DH5α中,扩大培养后测序鉴定。同时,运用ORF Finder、BLAST、Protparam、ProtScaleon Expasy、TMHMM Server v.2.0、SignalP4.1 Server、NetPhos3.1 Server、NetAcet1.0 Server、NetNGlyc1.0 Server、SOPMA、SWISS-MODEL、ABCpred、STRING等生物信息学分析工具对PPE7蛋白可能的相关生物学信息进行预测。结果显示,Rv0354c基因成功扩增,测序正确。基因全长426 bp,有2个开放阅读框,其编码蛋白为PPE7,由141个氨基酸组成,等电点为5.45,为疏水性蛋白,跨膜氨基酸数为0.59727,无信号肽、乙酰化位点和N-糖基化位点,有14个磷酸化位点;并且PPE7具有潜在的T、B细胞表位,与多种蛋白质存在相互作用关系,可能为抗结核疫苗的研发提供重要参考。The gene sequence encoding PPE7 protein was obtained from NCBI. The Rv0354c gene was cloned from H37Rv genome with designed primers, transformed into E. coli competent cell DH5α, sequenced and identified after expanded culture. In addition, the possible biological information related to PPE7 protein was predicted by ORF finder, Blast, ProtParam, ProtScaleon Exeasy, TMHMM Serverv.2.0, SignalP4.1 Server,NetPHOS3.1 Server, NetAcet1.0 Server, NetNGLYC1.0 Server, SOPMA, Swiss-Model, ABCpred, STRING and other tools. The Rv0354c gene was successfully amplified and sequenced. The full length of Rv0354c gene was 426 bp, with two open reading frames, encoding protein PPE7, which consists of 141 amino acids,isoelectric point is 5.45, and it is a hydrophobic protein. The number of transmembrane amino acids is 0.59727.There are 14 phosphorylation sites, without signal peptide, acetylation site and N-glycosylation site. PPE7 has potential T and B cell antigen epitopes and interacts with a variety of proteins, which may provide an important reference for the development of anti-tuberculosis vaccines.

关 键 词：PPE7蛋白 结核分枝杆菌 Rv0354c 生物信息学分析 

分 类 号：R378.911[医药卫生—病原生物学]