Construction of a recombinant plasmid for S.mutants luxS gene knock-out / 实用口腔医学杂志

ABSTRACT

Objective:To construct a recombination plasmid containing a kanamycin resistance gene,the upstream and downstream fragment of luxS of Streptococcus mutans so that luxS can be knock out by transforming the plasmid into S.mutans later.Methods:Kanamycin resistance gene,the upstream and downstream of luxS were cloned respectively by using plasmid pEGFP-N1 and DNA of Streptococcus mutans as template.Then the genes were ligated into Multiple Cloning Site(MCS) of vector pMD19-T in certain order and transformed into E.coli Competent Cells.Finally transformants were selected for resistance to kanamycin and ampicillin.Results:Kanamycin resistance gene and the upstream and downstream of luxS were successfully ligated into accurate enzyme digestion site of vector pMD19-T,and restriction digests analysis and sequencing result was correct.Conclusion:LuxS gene knock-out of Streptococcus Mutans recombinant plasmid is constructed and built a base of constructing Streptococcus Mutans luxS mutans in the future.