军事医学2017，Vol.41Issue(3)：209-212,221,5.DOI:10.7644/j.issn.1674-9960.2017.03.011
鼠疫耶尔森菌基因组无痕修饰方法的建立
Development of a high-efficient scarless genetic modification method for Yersinia pestis
摘要
Abstract
Objective To construct a technical platform for scarless gene modification of Yersinia pestis and to study the functions of its specific genes.Methods The resistance fragment, including upstream and downstream homologous arms of targeted regions, was reamplified by asymmetric PCR.The amplicons were introduced into Y.pestis harboring plasmid pKD46.With the induction of L-arabinose,the recombinant related enzymes: Exo, Beta and Gam, were expressed to guide the homologous recombination.A donor plasmid, pKSI-1, which carried the desired modification fragment flanking by I-SceⅠ recognition sites, was introduced into Y.pestis as the second step of λ-Red recombination with the help of pREDTKI.Results and Conclusion Two mutant strains:△waaA and waaA(△9nt), were successfully constructed for Y.pestis strain 201.Scarless modification introduces no extra modification to the genome, and it is ideal for comprehensive functional genomic studies.关键词
鼠疫菌/同源重组/基因编辑Key words
Yersinia pestis/homologous recombination/genome modification分类
生物科学引用本文复制引用
肖丽生,祁芝珍,吕瑞辰,宋凯,陈荣,王敏,吴海莲,赵海红,宋亚军..鼠疫耶尔森菌基因组无痕修饰方法的建立[J].军事医学,2017,41(3):209-212,221,5.基金项目
国家自然科学基金面上资助项目(31360034,31470138) （31360034,31470138）
