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中文摘要：
为探讨肠炎沙门菌非编码小RNA(non-coding small RNA)IsrE在转录后水平是否参与对fimA的调控作用,在大肠杆菌中构建并表达FimA重组蛋白,并对其培养条件进行优化,制备高纯度FimA重组蛋白。利用原核表达载体p ET-28a(+)构建出能表达fimA的重组质粒,并分别对IPTG浓度、诱导温度和诱导时间等条件进行优化,以提高目的蛋白的产量和增大其可溶性,最后经Ni-NTA亲和层析分离纯化FimA重组蛋白。通过酶切、测序和Western blot鉴定分析,成功构建并表达肠炎沙门菌I型菌毛主要亚单位FimA,蛋白分子质量约为19.3 kDa,且主要以包涵体的形式表达,经Ni-NTA亲和层析纯化后的重组蛋白在SDS-PAGE中显示单一条带。本研究构建、表达并获得纯度较高的重组蛋白FimA,为进一步研究和验证肠炎沙门菌非编码sRNA IsrE是否在转录后水平参与对fimA的调控奠定了基础。
英文摘要：
To investigate if the non-coding small RNA IsrE of Salmonella enteritidis was involved in regulating FimA at the post-transcriptional level, fimA gene was cloned in pET-28a(+) vector that was then transformed into E.coli BL21(DE3). The expression conditions of FimA were optimized and expressed FimA was purified using Ni2+-chelating chromatography. The recombinant FimA was visualized as a single clear band of a relative molecular weight at 19.3 kDa in SDS-PAGE. The availability of highly purified recombinant FimA provided the foundation and platform for further study on whether or not FimA acted as a candidate target gene for small non-coding RNA IsrE of Salmonellaenteritidis at the post-transcriptional level.
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Antimicrobial resistance, presence of integrons and biofilm formation of Salmonella Pullorum isolate
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