山西医科大学学报2023，Vol.54Issue(12)：1591-1599,9.DOI:10.13753/j.issn.1007-6611.2023.12.007
结核分枝杆菌GroEL2蛋白表达、纯化及生物信息学分析
Expression,purification,and bioinformatics analysis of Mycobacterium tuberculosis GroEL2 protein
摘要
Abstract
Objective To clone,express,and purify the GroEL2 protein of Mycobacterium tuberculosis strain H37Ra and perform its bioinformatics analysis.Methods GroEL2 gene was amplified with the genomic DNA of Mycobacterium tuberculosis strain H37Ra as the template by PCR,and then cloned into the pET-28a vector to construct a recombinant pET-28a-GroEL2 vector,which was transformed into BL21(DE3)strain and expressed in an isopropyl β-D-thiogalactoside(IPTG).The expression products were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE),the recombinant GroEL2 protein was purified by nickel-iminodiacetic acid(Ni-IDA)affinity chromatography column,and the expression efficacy of GroEL2 was identified by Western blot.Bioinformatics methods were used to predict the physicochemical properties,the protein structure,and the antigenic epitopes of GroEL2 protein.Results The recombinant pET-28a-GroEL2 expression vector was successfully constructed and the GroEL2 protein was expressed in soluble form in E.coli.The recombinant GroEL2 protein was purified by Ni-IDA affinity chromatography column with a purity of over 90％.Bioinformatics analysis showed that it could interact with a variety of proteins and there were several potential T and B cell antigenic epitopes.Conclusion The recombinant GroEL2 protein has good immunoreactivity and can be used as a new target for the development of novel vaccines and the diagnosis for tuberculosis.关键词
结核分枝杆菌/GroEL2蛋白/原核表达/蛋白纯化/生物信息学Key words
Mycobacterium tuberculosis/GroEL2 protein/prokaiyotic expression/protein purification/bioinformatics分类
医药卫生引用本文复制引用
宋亚敏,魏婧,郭方正,李柏青,许涛,汪洪涛..结核分枝杆菌GroEL2蛋白表达、纯化及生物信息学分析[J].山西医科大学学报,2023,54(12):1591-1599,9.基金项目
安徽省自然科学基金项目(1908085MH252,2008085QH405) （1908085MH252,2008085QH405）
慢性疾病免疫学基础与临床安徽省重点实验室开放课题基金项目(KLICD-2002-Z3) （KLICD-2002-Z3）
呼吸系病临床基础安徽省重点实验室开放课题基金项目(HX2022-Z02) （HX2022-Z02）
蚌埠医学院"512人才培育计划"项目(by51201309) （by51201309）
蚌埠医学院研究生科研创新计划项目(Byy-cx22013,Byycx23078) （Byy-cx22013,Byycx23078）
