河南科技大学学报（医学版）2017，Vol.35Issue(1)：22-24,3.DOI:10.15926/j.cnki.issn1672-688x.2017.01.007
结核分枝杆菌H37Rv蛋白Rv2364c的表达纯化与生物信息学分析
Expression, Purification and Bioinformatics Analysis of Rv2364c from Mycobacterium Tuberculosis H37Rv
摘要
Abstract
Objective To obtain Rv2364c of Mycobacterium tuberculosis H37Rv expressed in E.coli and predict its bioinformatics characters for the further development of anti-Mycobacterium tuberculosis drugs.Methods The target DNA fragment of the Rv2364c gene was amplified by PCR and cloned into the prokaryotic expression vector pET-28a.Then the recombinant vector (pET-28a-Rv2364c) was transformed into E.coli BL21(DE3).The expression of the His-Tag fusion protein was induced with isopropyl-β-D-thiogalactoside(IPTG).Finally, we purified the protein through affinity chromatography.We also used bioinformatic methods to analyze and predict this protein.Results Double restriction enzyme analysis and sequencing technique proved that recombinant plasmid was constructed correctly, and the expression of the fusion protein in E.coli BL21(DE3) was detectable by SDS-PAGE.The target protein was gained after purified (purity>90%) using the affinity chromatographic column.Bioinformatics analysis revealed that Rv2364c was a stable and acidic protein;α-helix and random coil were the main components of its secondary structure;The main function domain types were Era domain and KH domain.ConclusionRv2364c had been successfully cloned, expressed and purified, the domain was predicted by bioinformatics method,which would be useful for further research on Rv2364c in the genesis and development of tuberculosis.关键词
结核分枝杆菌H37Rv/Rv2364c/转染/生物信息分析Key words
mycobacterium tuberculosis H37Rv/Rv2364c/transfection/bioinformatics分类
生物科学引用本文复制引用
徐宛玲,付陈增..结核分枝杆菌H37Rv蛋白Rv2364c的表达纯化与生物信息学分析[J].河南科技大学学报（医学版）,2017,35(1):22-24,3.基金项目
漯河医学高等专科学校基金(2014-S-LMC07) （2014-S-LMC07）
