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首页> 外文期刊>Iranian journal of science and technology >Recombinant Human SCARB2 Expressed in Escherichia coli and its Potential in Enterovirus 71 Blockage
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Recombinant Human SCARB2 Expressed in Escherichia coli and its Potential in Enterovirus 71 Blockage

机译:在大肠杆菌中表达的重组人类围巾及其在肠道病毒71堵塞的潜力

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摘要

Hand, foot and mouth disease is a common viral infectious disease caused by enteroviruses, including coxsackie A16 (CVA16) and enterovirus 71 (EV71). HFMD can cause severe symptoms in children which can be fatal. Human scavenger receptor class B member 2 (SCARB2) is a cellular receptor for EV71 and CVA16, providing a potential approach for preventing EV71 infection and transmission. In this present study, we constructed and assessed the potential of recombinant SCARB2, using E. coli expression system. To generate this construct, scarb2 gene was cloned into pET22b vector and expressed in E. coli BL21 (DE3). The expression of SCARB2 was induced by 0.1 mM IPTG and analyzed using SDS-PAGE, followed by Western blot. Expressed SCARB2 was in inclusion bodies and refolded to obtain the soluble form with recovery efficacy of 100%. This recombinant protein was then validated for binding with EV71 via indirect ELISA in two different pHs (7.4 and 5.5), which partially revealed the mechanism of virus-receptor interaction. These results envisaged potential applications for utilizing recombinant SCARB2 in preventing the virus transmission.
机译:手足口病是由肠道病毒引起,包括手足口A16(CVA16)和肠道病毒71(EV71)的公共病毒感染性疾病。手足口病在儿童有致命的危险引起严重的症状。人类清道夫受体B类成员2(SCARB2)为EV71和CVA16的细胞受体,从而提供用于防止EV71感染和传播的潜在方法。在此研究中,我们构建和评估重组SCARB2的潜力,使用大肠杆菌表达系统。为了产生该构建体,SCARB2基因克隆入的pET22b载体中并在大肠杆菌BL21(DE3)中表达。 SCARB2的诱导表达0.1mM的IPTG和使用SDS-PAGE,随后通过Western印迹进行分析。表达SCARB2在包涵体和复性以获得具有100%的回收率功效可溶形式。然后该重组蛋白质进行了验证为在两个不同的pH值(7.4和5.5),其部分地揭示病毒 - 受体相互作用的机制与EV71通过间接ELISA结合。这些结果设想潜在应用利用重组SCARB2防止病毒传播。

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