本文選題：大腸桿菌 + 重組蛋白�。� 參考：《口腔醫(yī)學(xué)研究》2017年02期

【摘要】：目的:探究誘導(dǎo)重組GroEL(牙周炎相關(guān)動(dòng)脈粥樣硬化疫苗)的最佳IPTG濃度,并分析IPTG濃度影響重組GroEL表達(dá)量的原因。方法:構(gòu)建GroEL的大腸桿菌(Escherichia Coli,E.coli)表達(dá)體系,使用不同濃度IPTG(0,10,20,30,50,100μmol/L)誘導(dǎo)構(gòu)建的E.coli(GroEL-E.coli),運(yùn)用SDS-PAGE及相關(guān)軟件定量分析GroEL表達(dá)量;透射電子顯微鏡及A值測(cè)定法分別檢測(cè)IPTG濃度對(duì)GroEL-E.coli內(nèi)部包涵體(IB)形成及GroEL-E.coli生長(zhǎng)的影響。結(jié)果:重組GroEL主要以可溶性蛋白的形式存在。IPTG的加入可增加GroEL表達(dá)量,當(dāng)IPTG濃度為30μmol/L時(shí)GroEL表達(dá)量最多。IPTG可促進(jìn)IB形成并抑制GroEL-E.coli生長(zhǎng),且影響效果隨IPTG濃度增加而增強(qiáng)。結(jié)論:IPTG濃度影響GroEL-E.coli中重組GroEL的表達(dá),最佳IPTG濃度為30μmol/L;IPTG可通過誘導(dǎo)IB形成及抑制細(xì)菌增殖影響GroEL表達(dá)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.
【作者單位】： 南京大學(xué)醫(yī)學(xué)院附屬口腔醫(yī)院南京市口腔醫(yī)院牙周病科;
【基金】：國(guó)家自然科學(xué)基金(編號(hào):81271155;51472115) 南京市科技發(fā)展計(jì)劃項(xiàng)目(編號(hào):201402036) 南京市醫(yī)學(xué)科技發(fā)展項(xiàng)目(編號(hào):YKK14107)
【分類號(hào)】：R781.4

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