Ianov S N, Darmov I V, Marakulin I V
Mol Gen Mikrobiol Virusol. 1993 Sep-Oct(5):16-9.
The recombinant plasmid pKC47M has been constructed on the base of a mini-Mu-phage and the replicon of plasmid pBR328. The plasmid was constructed for in vivo insertional mutagenesis in bacteria. Use of the plasmid in vivo alleviates the possibility of secondary transposition of the inserted sequences and enrichment of multiple-site mutations. Efficient mutagenesis caused by plasmid pKC47M was tested by induction auxotrophic mutations in Escherichia coli, Yersinia enterocolitica, Yersinia pseudotuberculosis.
重组质粒pKC47M是在微型Mu噬菌体和质粒pBR328的复制子基础上构建的。该质粒用于细菌体内插入诱变。在体内使用该质粒可减少插入序列二次转座的可能性以及多位点突变的富集。通过在大肠杆菌、小肠结肠炎耶尔森菌、假结核耶尔森菌中诱导营养缺陷型突变来测试质粒pKC47M引起的高效诱变。