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狐肺炎大腸桿菌HtrA蛋白原核表達(dá)及多抗血清制備

發(fā)布時(shí)間:2019-06-24 14:15
【摘要】:利用PCR方法從銀狐大腸桿菌基因組中克隆出htra基因,測(cè)序無(wú)誤后將其構(gòu)建到pET32a原核表達(dá)載體上,重組載體轉(zhuǎn)化BL21大腸桿菌誘導(dǎo)重組蛋白表達(dá),通過(guò)SDS-PAGE和Western blot方法驗(yàn)證重組蛋白得到很好表達(dá);用親和層析方法純化蛋白,純化的蛋白免疫小鼠,所得的多抗血清能夠特異性識(shí)別大腸桿菌HtrA蛋白,為研究HtrA蛋白的作用機(jī)制奠定基礎(chǔ)。
[Abstract]:The hra gene was cloned from the genome of the E. coli. The recombinant vector was transformed into E. coli to induce the expression of the recombinant protein, and the recombinant protein was expressed by SDS-PAGE and Western blot. The purified protein was purified by affinity chromatography, and the purified protein was used to immunize the mouse. The obtained polyclonal antiserum can specifically identify the E. coli HtrA protein and lay the foundation for studying the mechanism of the action of the HtrA protein.
【作者單位】: 河北科技師范學(xué)院河北省預(yù)防獸醫(yī)學(xué)重點(diǎn)實(shí)驗(yàn)室;秦皇島第二醫(yī)院;
【基金】:國(guó)家“星火計(jì)劃”資助項(xiàng)目(2015GA620002) 河北省科技廳“科技支撐計(jì)劃”資助項(xiàng)目(14826613D) 河北科技師范學(xué)院博士啟動(dòng)基金資助項(xiàng)目(2015YB002) 河北省高等學(xué)?茖W(xué)技術(shù)研究資助項(xiàng)目(ZD2016067) 秦皇島市農(nóng)業(yè)科學(xué)研究院資助項(xiàng)目(2014-04)
【分類號(hào)】:S852.61
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本文編號(hào):2505124

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