金黄葡萄球菌肠毒素C3基因的克隆及原核表达

目的克隆并原核表达金黄葡萄球菌肠毒素C3(SEC3)基因。方法以金黄葡萄球菌基因组DNA为模板,扩增SEC3基因中编码成熟蛋白的基因片段,克隆至pET-32a(+)表达载体,构建重组表达质粒SEC3-pET-32a,转化E.coli BL21(DE3),IPTG诱导表达。表达产物经Ni-NTA亲和层析纯化后,进行SDS-PAGE和Western blot分析。结果经PCR扩增获得720bp的SEC3基因;重组表达质粒构建正确;表达产物的相对分子质量约为43000,为可溶性表达;纯化的重组蛋白具有良好的反应原性。结论已成功克隆并原核表达了SEC3基因,为单克隆抗体和诊断试剂的制备及SEC3抗毒素的研制提供了材料。

Cloning and Prokaryotic Expression of Stapylococcus aureus Enterotoxin C3 Gene

Objective To clone Stapylococcus aureus enterotoxin C3(SEC3)gene and express in prokaryotic cells.Methods The SEC3 gene fragment encoding mature protein was amplified by PCR using the genomic DNA of Stapylococcus aureus as a template,and cloned into vector pET-32a(+).The recombinant plasmid SEC3-pET-32a was transformed to E.coli BL21(DE3)for expression under induction of IPTG.The expressed product was purified by Ni-NTA affinity chromatography and identified by SDSPAGE and Western blot.Results The SEC3 gene...