伤寒沙门菌sufC基因缺陷变异株的制备

Construction of sufC gene deleted mutant in salmonella enterica serovar typhi

目的为深入研究sufC基因在伤寒沙门菌中的功能作用,制备sufC基因缺陷变异株。方法根据GeneBank公布的伤寒沙门菌sufC基因序列,设计sufC缺失用PCR特异性引物,制备缺失sufC基因的同源性核苷酸片段,连接自杀质粒后导入伤寒沙门菌野生株,诱导同源重组,重组菌经PCR观察及序列分析鉴定,将完全重组稳定的相应菌株作为伤寒沙门菌sufC基因缺陷变异株,并经测序分析加以确定。结果 PCR及序列分析证实,缺陷变异株的sufC基因缺失495个碱基。结论伤寒沙门菌sufC基因缺陷株构建成功,为进一步研究其在伤寒沙门菌中的功能作用奠定了基础。

Objective For investigating the funcation of sufC gene,the deletion mutant of the sufC gene was constructed in Salmonella enterica serovar Typhi（S.typhi）.Methods According to sufC gene sequence from the Gene Bank,designed sufC deletions using PCR specific primer.Constructed deletions of the sufC gene homology in nucleotide fragment,and connected the suicide plasmid.The suicide plasmid including sufC gene homology in nucleotide fragment which was transferred into S.typhi wild strains.It was induction of homologous recombination.The recombination was valided by PCR and sequencing analysis.The completely stable recombinant strains were sufC gene defective mutant of S.typhi.Results PCR and sequencing analysis confirmed that the sufC gene mutant was deleted 495bp.Conclusions The sufC gene deleted mutant of S.enterica serovar Typhi was generated successfully which was a foundation to study the function of the sufC gene in S.enterica serovar Typhi.