Willis D K, Satin L H, Clark A J
J Bacteriol. 1985 Jun;162(3):1166-72. doi: 10.1128/jb.162.3.1166-1172.1985.
Using pBR322 as a vector, we cloned a 5.95-kilobase fragment of the Rac prophage together with 1.70 kilobases of a flanking Escherichia coli chromosome sequence. The resulting plasmid (pRAC1) was unable to suppress the mitomycin and UV sensitivity and recombination deficiency of a recB21 recC22 strain. Five spontaneous mitomycin-resistant derivatives contained deletion mutant plasmids. These plasmids also suppressed the UV sensitivity and recombination deficiency of their recB21 recC22 hosts. All five deletions were contained within a 2.45-kilobase EcoRI-to-HindIII segment of the plasmid. By substituting the corresponding 2.45-kilobase EcoRI-toHindIII fragments of Rac prophage isolated from sbcA+, sbcA6, and sbcA23 strains for the shortened segment of one of the deletion mutant plasmids, we were able to show that sbcA mutations map in this region. Also in this region is the site (or closely linked sites) at which previous studies had shown that insertion of Tn5 and IS50 leads to suppression of recB21 recC22. The sequence in this region that must be altered or circumvented to allow suppression is discussed. Also presented are data correlating the expression of nuclease activity with the degree of suppression.
以pBR322作为载体,我们克隆了Rac原噬菌体的一个5.95千碱基片段以及1.70千碱基的侧翼大肠杆菌染色体序列。所得质粒(pRAC1)无法抑制recB21 recC22菌株对丝裂霉素和紫外线的敏感性以及重组缺陷。五个自发的丝裂霉素抗性衍生物含有缺失突变体质粒。这些质粒也抑制了其recB21 recC22宿主对紫外线的敏感性和重组缺陷。所有五个缺失都包含在质粒的一个2.45千碱基的EcoRI至HindIII片段内。通过用从sbcA +、sbcA6和sbcA23菌株分离的Rac原噬菌体的相应2.45千碱基EcoRI至HindIII片段替换其中一个缺失突变体质粒的缩短片段,我们能够证明sbcA突变位于该区域。之前的研究表明,Tn5和IS50的插入会导致recB21 recC22的抑制,该区域也是插入位点(或紧密连锁的位点)。讨论了该区域中必须改变或规避以实现抑制的序列。还展示了将核酸酶活性表达与抑制程度相关联的数据。