IPTG浓度对大肠杆菌中重组GroEL表达的影响

发布时间：2018-04-10 17:19

  本文选题：大肠杆菌 + 重组蛋白　； 参考：《口腔医学研究》2017年02期

【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。
[Abstract]:Aim: to investigate the optimal IPTG concentration of recombinant Groelle (periodontitis associated atherosclerotic vaccine) and analyze the reasons why IPTG concentration affects the expression of recombinant GroEL.The effects of IPTG concentration on the formation of GroEL-E.coli inclusion bodies and the growth of GroEL-E.coli were detected by transmission electron microscope and A value method respectively.Results: the expression of GroEL was increased by the addition of. IPTG in the form of soluble protein. When the concentration of IPTG was 30 渭 mol/L, the expression of GroEL was the most. IPTG could promote the formation of IB and inhibit the growth of GroEL-E.coli, and the effect increased with the increase of IPTG concentration.Conclusion the expression of recombinant GroEL in GroEL-E.coli was affected by the concentration of 10% IPTG. The optimal concentration of IPTG was 30 渭 mol / L IPTG, which could affect the expression of GroEL by inducing IB formation and inhibiting bacterial proliferation.
【作者单位】： 南京大学医学院附属口腔医院南京市口腔医院牙周病科;
【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)
【分类号】：R781.4

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