MBP-CsgAⅢ融合蛋白的表达和纯化

Expression and purification of MBP-CsgAⅢ fusion protein

目的构建CsgA亚单位蛋白的原核表达系统,获得纯化的MBPCsgAⅢ融合蛋白,为进一步研究CsgA的生物学特性和致病机理奠定基础。方法用PCR的方法从大肠杆菌野生株MC4100株中扩增出csgA基因,连接到pMALp2质粒上,构建pZBaⅢ重组质粒。将重组质粒转入XL1Blue菌中用IPTG诱导表达,用AmyloseResin纯化系统纯化。结果成功构建了MBPCsgAⅢ融合蛋白的原核表达系统,获得大量纯化的MBPCsgAⅢ融合蛋白。结论成功构建了MBPCsgAⅢ融合蛋白的原核表达系统,应用该系统能有效表达MBPCsgAⅢ融合蛋白,为进一步研究CsgA的生物学特性和致病机理奠定了基础。

Objective To express and purify the MBP-CsgAⅢ fusion protein for studying the biogenesis and pathogenesis of bacterial fimbriae curli in E. coli. Methods The csgA gene was amplified by PCR from E, coli MC4100 strain, and then inserted into plasmid pMAL-p2, resulting in pZS-aⅢ, The pZB-aⅢ was transformed into XL1-Blue strain, The XL1 -Blue with pZB-aⅢ strain was induced by IPTG to over-express MBP-CsgA fusion protein, which was affinity-purified by using amylose resin purification system. Results The MBP-CsgAⅢ fusion protein expression system was established successfully. The MBP-CsgA m fusion protein was successfully over-expressed. Conclusion The MBP-CsgAm fusion protein is expressed and purified by using the constructed expression system, which will benefits the research on biogenesis and pathogenesis of curli in E, coli.