Fouts K E, Wasie-Gilbert T, Willis D K, Clark A J, Barbour S D
J Bacteriol. 1983 Nov;156(2):718-26. doi: 10.1128/jb.156.2.718-726.1983.
Fourteen mitomycin-resistant revertants of a recB21 recC22 strain were isolated after Tn5 mutagenesis. Eight of the mutations (type I) were essentially inseparable from aphA+ (Kanr) of Tn5; six (type II) were not. We hypothesize that the former are Tn5 and that the latter are IS50 insertions. Because of their phenotypic similarity to sbcA and sbcB mutations, which also suppress recB21 recC22, we have called them sbc mutations. sbc-lll::Tn5 was cotransducible with nirR and has thereby been located at position 29.8 on the Escherichia coli map in the vicinity of the Rac prophage and sbcA mutations. A recB21 recC22 sbc-lll::Tn5 strain was subjected to Tn10 mutagenesis, and a mitomycin- and UV-sensitive mutant was isolated. tet+ of Tn10 was 85% cotransducible with aphA+ of Tn5, locating these two transposons 0.1 map unit apart. A three-point cross located the Tn10 mutation at position 29.7. We hypothesize that the Tn10 insertion is located in recE and that the Tn5 and IS50 insertions activate expression of this gene. sbc-lll::Tn5 was found to be cis acting and dominant to its wild-type allele as were two sbcA mutations (sbcA1 and sbcA6). Five other type I and type II insertion mutations were dominant to their wild-type alleles. We hypothesize that the sbc insertion and sbcA mutations affect transcription regulation of recE and discuss the possibility that they do so differently.
在Tn5诱变后,分离出了recB21 recC22菌株的14个丝裂霉素抗性回复突变体。其中8个突变(I型)与Tn5的aphA +(卡那霉素抗性)基本不可分离;6个(II型)则不是。我们推测前者是Tn5插入,后者是IS50插入。由于它们在表型上与sbcA和sbcB突变相似,后者也能抑制recB21 recC22,所以我们将它们称为sbc突变。sbc-III::Tn5与nirR共转导,因此定位于大肠杆菌染色体图谱上29.8位置,靠近Rac原噬菌体和sbcA突变。对recB21 recC22 sbc-III::Tn5菌株进行Tn10诱变,分离出一个对丝裂霉素和紫外线敏感的突变体。Tn10的tet +与Tn5的aphA +共转导率为85%,表明这两个转座子相距0.1个图谱单位。三点杂交将Tn10突变定位在29.7位置。我们推测Tn10插入位于recE中,并且Tn5和IS50插入激活了该基因的表达。发现sbc-III::Tn5与两个sbcA突变(sbcA1和sbcA6)一样,是顺式作用且对其野生型等位基因呈显性。其他5个I型和II型插入突变对其野生型等位基因也是显性的。我们推测sbc插入和sbcA突变影响recE的转录调控,并讨论了它们以不同方式调控的可能性。