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- 2017
IPTG浓度对大肠杆菌中重组GroEL表达的影响
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Abstract:
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(0,10,20,30,50,100 μmol/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30 μmol/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30 μmol/L;IPTG可通过诱导IB形成及抑制细菌增殖影响GroEL表达量
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