IPTG浓度对大肠杆菌中重组GroEL表达的影响  

Effect of IPTG Concentrations on Recombinant Protein GroEL Production in Escherichia Coli

作　　者：肖玲玲[1] 杨洁[1] 张瑞[1] 陈小凤[1] 刘玉[1] 吴娟[1] 苗雷英[1] 孙卫斌[1] 

机构地区：[1]南京大学医学院附属口腔医院,南京市口腔医院牙周病科,江苏南京210008

出　　处：《口腔医学研究》2017年第2期140-144,共5页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:81271155;51472115);南京市科技发展计划项目(编号:201402036);南京市医学科技发展项目(编号:YKK14107)

摘　　要：目的:探究诱导重组GroEL(牙周炎相关动脉粥样硬化疫苗)的最佳IPTG浓度,并分析IPTG浓度影响重组GroEL表达量的原因。方法:构建GroEL的大肠杆菌(Escherichia Coli,E.coli)表达体系,使用不同浓度IPTG(0,10,20,30,50,100μmol/L)诱导构建的E.coli(GroEL-E.coli),运用SDS-PAGE及相关软件定量分析GroEL表达量;透射电子显微镜及A值测定法分别检测IPTG浓度对GroEL-E.coli内部包涵体(IB)形成及GroEL-E.coli生长的影响。结果:重组GroEL主要以可溶性蛋白的形式存在。IPTG的加入可增加GroEL表达量,当IPTG浓度为30μmol/L时GroEL表达量最多。IPTG可促进IB形成并抑制GroEL-E.coli生长,且影响效果随IPTG浓度增加而增强。结论:IPTG浓度影响GroEL-E.coli中重组GroEL的表达,最佳IPTG浓度为30μmol/L;IPTG可通过诱导IB形成及抑制细菌增殖影响GroEL表达量。Objective：To explore the effect of IPTG concentrations on recombinant GroEL production in Escherichia Coli（E.coli）and analyze the reasons.Methods：The E.coli expression system for GroEL was constructed and induced by different IPTG concentrations（0,10,20,30,50,100μmol/L）for GroEL expression,and the SDS-PAGE and gel analysis software were used to determine the amount of expressed GroEL.The transmission electron microscopy（TEM）and OD-measuring were used to detect IB formation and bacterial growth.Results：GroEL mainly exists in the soluble form.The addition of IPTG can obviously increase the GroEL expression,and 30μmol/L IPTG leaded to the most GroEL expression.Conclusion：The recombinant GroEL expression in E.coli differed among IPTG concentrations,and the optimal IPTG concentration was 30μmol/L.The main reasons for IPTG effect on GroEL yield were IB formation and growth inhibition.

关 键 词：大肠杆菌 重组蛋白 Isopropyl-β-D-thiogalactopyranoside 

分 类 号：R781.4[医药卫生—口腔医学]