Oden K L, DeVeaux L C, Vibat C R, Cronan J E, Gennis R B
Department of Biochemistry, University of Illinois, Urbana 61801.
Gene. 1990 Nov 30;96(1):29-36. doi: 10.1016/0378-1119(90)90337-q.
A number of gene replacements at different loci were constructed using covalently closed circular (ccc) plasmid DNA in the recB21 recC22 sbcB15 sbcC201 mutant of Escherichia coli (JC7623). Selected constructs representing deletions and insertion mutations formed from double-crossover events involving the ccc plasmid molecules and the genome were confirmed by Southern blots, and the frequency of double-crossover events was evaluated. It is reported that such mutants may be constructed without linearizing plasmid DNA, as described previously.
利用共价闭合环状（ccc）质粒DNA，在大肠杆菌（JC7623）的recB21 recC22 sbcB15 sbcC201突变体中构建了多个不同位点的基因替换。通过Southern杂交证实了由涉及ccc质粒分子和基因组的双交换事件形成的代表缺失和插入突变的选定构建体，并评估了双交换事件的频率。据报道，如前所述，此类突变体可以在不使质粒DNA线性化的情况下构建。
