Villani G, Canosi U, Fortunato A, Mazza G, Polsinelli M, Falaschi A
Nucleic Acids Res. 1974 Mar;1(3):461-77. doi: 10.1093/nar/1.3.461.
We have isolated a mutant of Bacillussubtilis deficient in DNA polymerase I, denominated polA42, which shows a reduced ability to repair the damage to DNA by UV radiation, MMS and mitomycin C;the ability to perform recombination is not appreciably impaired.DEAE cellulose chromatography allows the separation of polymerases I and II from the parental strain;a simple procedure is also described which allows to separate rapidly the polymerases II and III of the mutant strain. The three separated polymerases have similar catalytic properties but they can be distinguished for their sensitivity to inhibitors: PCMB inhibits polymerases II and III but not polymerase I; HPUra inhibits only polymerase III. All three enzymes are unaffected by nalidixate. The DNA synthesis occurring in cells of the polA42 strain permeabilized with toluene is inhibited by nalidixate, whereas the synthesis occurring in polA(+) toluenized cells is unaffected by the drug. The polA gene has been mapped by transduction and localized between the phe(12) and argA(3) genes.
我们已经分离出一株枯草芽孢杆菌的DNA聚合酶I缺陷型突变体,命名为polA42,该突变体对紫外线、甲基磺酸甲酯和丝裂霉素C造成的DNA损伤的修复能力降低;但其进行重组的能力并未明显受损。通过DEAE纤维素层析可以从亲本菌株中分离出聚合酶I和II;还描述了一种简单的方法,可快速分离突变菌株的聚合酶II和III。分离出的三种聚合酶具有相似的催化特性,但可根据它们对抑制剂的敏感性加以区分:对氯汞苯甲酸抑制聚合酶II和III,但不抑制聚合酶I;羟基脲仅抑制聚合酶III。这三种酶均不受萘啶酸的影响。用甲苯透化处理的polA42菌株细胞中的DNA合成受到萘啶酸的抑制,而经甲苯处理的polA(+)细胞中的合成不受该药物影响。通过转导对polA基因进行了定位,其位于phe(12)和argA(3)基因之间。