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目的原核表达并纯化沙门氏菌毒性相关蛋白STM1697,为其功能研究奠定基础。方法以鼠伤寒沙门氏菌(ATCC14028)基因组为模板,PCR扩增STM1697基因,双酶切后连接到原核表达载体pGL01中。将阳性重组子转化入大肠埃希菌BL21(DE3)以IPTG诱导,表达产物经镍离子亲和柱进行纯化,并用SDS-PAGE分析鉴定蛋白及纯度。结果成功将STM1697基因克隆到原核表达载体pGL01中。STM1697蛋白在大肠埃希菌BL21(DE3)以可溶形式表达,经镍离子亲和柱纯化后得到的蛋白纯度>95%。结论伤寒沙门氏菌STM1697蛋白在大肠埃希菌中可稳定表达,为其功能研究奠定了基础。
Objective To express and purify Salmonella virulence-associated protein STM1697 in prokaryotic cells and lay the foundation for its function. Methods The genomic DNA of Salmonella typhimurium (ATCC14028) was used as template to amplify the STM1697 gene by PCR and ligated into prokaryotic expression vector pGL01. The positive recombinant was transformed into Escherichia coli BL21 (DE3) induced by IPTG. The expressed product was purified by nickel ion affinity column, and the protein and purity were identified by SDS-PAGE analysis. Results The STM1697 gene was successfully cloned into prokaryotic expression vector pGL01. The STM1697 protein was expressed in soluble form in Escherichia coli BL21 (DE3), and the purity of the purified protein was> 95% after purified by nickel ion affinity column. Conclusion Salmonella typhimurium STM1697 protein is stably expressed in Escherichia coli, which lays the foundation for its function research.