Manly K F, Smoler D F, Bromfeld E, Baltimore D
J Virol. 1971 Jan;7(1):106-11. doi: 10.1128/JVI.7.1.106-111.1971.
The in vitro product of mouse leukemia virus deoxyribonucleic acid (DNA) polymerase can be separated into two fractions by sedimentation in sucrose gradients. These two fractions were analyzed for their content of single-stranded DNA, double-stranded DNA, and DNA-ribonucleic acid (RNA) hybrid by (i) digestion with enzymes of known specificity and (ii) equilibrium centrifugation in Cs(2)SO(4) gradients. The major fraction early in the reaction contained equal amounts of single-stranded DNA and DNA-RNA hybrid and little double-stranded DNA. The major fraction after extensive synthesis contained equal amounts of single-and double-stranded DNA and little hybrid. In the presence of actinomycin D, the predominant product was single-stranded DNA. To account for these various forms of DNA, we postulate the following model: the first DNA synthesis occurs in a replicative complex containing growing DNA molecules attached to an RNA molecule. Each DNA molecule is displaced as single-stranded DNA by the synthesis of the following DNA strand, and the single-stranded DNA is copied to form double-stranded DNA either before or after release of the single strand from the RNA. Actinomycin blocks this conversion of single-to double-stranded DNA.
小鼠白血病病毒脱氧核糖核酸(DNA)聚合酶的体外产物可通过在蔗糖梯度中沉降分离为两个组分。通过以下方法分析这两个组分的单链DNA、双链DNA和DNA - 核糖核酸(RNA)杂交体含量:(i)用具有已知特异性的酶消化,以及(ii)在Cs₂SO₄梯度中进行平衡离心。反应早期的主要组分含有等量的单链DNA和DNA - RNA杂交体,双链DNA很少。大量合成后的主要组分含有等量的单链和双链DNA,杂交体很少。在放线菌素D存在下,主要产物是单链DNA。为了解释这些不同形式的DNA,我们提出以下模型:首次DNA合成发生在一个复制复合物中,该复合物包含附着于RNA分子的正在生长的DNA分子。随着下一条DNA链的合成,每个DNA分子作为单链DNA被置换,单链DNA在从RNA释放单链之前或之后被复制形成双链DNA。放线菌素阻止了单链DNA向双链DNA的这种转化。