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目的克隆伤寒沙门菌菌毛操纵子stg基因,在大肠埃希菌中表达;了解伤寒沙门菌stg与人上皮细胞的相互作用。方法根据大肠埃希菌保守基因序列设计引物,克隆stg成熟肽编码序列,连接到原核表达载体pET32a,构建重组表达质粒,转化至大肠埃希菌BL21(DE3)中并观察与上皮细胞的粘附特性。结果成功克隆stg基因全长核苷酸序列,构建了pET32a/stg原核表达重组质粒。pET32a/stg重组质粒转化大肠埃希菌能较多出现在上皮细胞表面。结论成功克隆并表达了伤寒沙门菌菌毛操纵子stg基因,stg基因能促进沙门菌对上皮细胞的粘附,为进一步了解伤寒沙门菌菌毛操纵子stg的特性与功能奠定了基础。
Objective To clone stg gene of Salmonella typhi pilus operon and express it in Escherichia coli. To understand the interaction between Salmonella typhi stg and human epithelial cells. Methods According to the conservative gene sequence of Escherichia coli, primers were designed. The stg mature peptide sequence was cloned and inserted into the prokaryotic expression vector pET32a. The recombinant plasmid was transformed into Escherichia coli BL21 (DE3) and observed the adhesion with epithelial cells characteristic. Results The full-length nucleotide sequence of stg gene was successfully cloned and the prokaryotic recombinant plasmid pET32a / stg was constructed. Escherichia coli transformed with pET32a / stg recombinant plasmid can appear more on the surface of epithelial cells. Conclusion Stg gene of Salmonella typhimurium was successfully cloned and expressed. The stg gene could promote the adherence of Salmonella to epithelial cells, which lays the foundation for further understanding of the characteristics and functions of Salmonella typhimurium stoger.