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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 To further study the functional role of sufC gene in Salmonella typhi, a mutant strain of sufC gene was prepared. Methods According to the sequence of sufC gene of Salmonella typhi isolated from GeneBank, homologous recombination primers for sufC gene were designed to amplify the nucleotide sequence of sufC gene. The sufC gene was inserted into wild strain of Salmonella typhi to induce homologous recombination. The recombinant strains were identified by PCR and sequence analysis. The completely recombinant stable strains were identified as defective mutants of sufC gene of Salmonella typhi, and sequenced. Results PCR and sequence analysis confirmed that the defective mutant sufC gene deletion of 495 bases. Conclusion SufC gene of S. typhi was successfully constructed, which laid the foundation for the further study of its function in Salmonella typhi.