Lee M, Miller R C
J Virol. 1974 Nov;14(5):1040-8. doi: 10.1128/JVI.14.5.1040-1048.1974.
The role of T7-induced exonuclease (gene 6) in molecular recombination was studied by examining the fate of parental DNA during parental-to-progeny recombination. The method used was to compare infections with T7(+), T7am-6-233 (am gene 6), or T7ts6-136 (ts gene 6) under permissive and nonpermissive conditions. CsCl density gradient analysis of replicative DNA indicated that T7 exonuclease is necessary for recombination to occur, i.e., in the absence of the exonuclease the parental DNA replicated continuously as a hybrid molecule and did not recombine. Further studies under conditions where replicative DNA was denatured and analyzed by CsCl density gradient centrifugation indicated that the exonuclease is also needed for a limited amount of covalent repair of recombinants. A repair function for the T7-induced exonuclease is also suggested by results obtained from alkaline sucrose gradient analysis of replicative DNA. Under conditions nonpermissive for the exonuclease, discontinuities in the DNA accumulated during infection by T7am6-233 or by T7ts6-136.
通过检查亲代 DNA 在亲代到子代重组过程中的命运,研究了 T7 诱导的核酸外切酶(基因 6)在分子重组中的作用。所采用的方法是在允许和非允许条件下比较用 T7(+)、T7am - 6 - 233(基因 6 突变体)或 T7ts6 - 136(基因 6 温度敏感突变体)进行的感染。对复制性 DNA 的 CsCl 密度梯度分析表明,T7 核酸外切酶是重组发生所必需的,即,在没有核酸外切酶的情况下,亲代 DNA 作为杂交分子连续复制且不发生重组。在复制性 DNA 变性并通过 CsCl 密度梯度离心分析的条件下进行的进一步研究表明,核酸外切酶对于重组体的有限共价修复也是必需的。对复制性 DNA 的碱性蔗糖梯度分析结果也表明 T7 诱导的核酸外切酶具有修复功能。在核酸外切酶非允许的条件下,T7am6 - 233 或 T7ts6 - 136 感染期间 DNA 中积累了不连续性。