Morrison D A, Guild W R
J Bacteriol. 1972 Dec;112(3):1157-68. doi: 10.1128/jb.112.3.1157-1168.1972.
The fate of label introduced as donor deoxyribonucleic acid (DNA) into competent cells of Diplococcus pneumoniae was determined immediately after entry at 25 C, as a function of the size of the donor DNA. Part of the label is found to be acid soluble, part has been incorporated into chromosomal DNA, apparently through reincorporation of degraded donor DNA, and part is found in single strands of length smaller than that of the input donor DNA strands. The last fraction apparently constitutes the precursor for integration of intact donor genetic markers and is referred to as the intact fraction. For large donor DNA the intact fraction contains over 80% of the total intracellular label, but the median strand length has been reduced to 2.2 x 10(6) daltons. For small donor molecules (1 x 10(5) to 6 x 10(5) daltons per strand) the fraction intact increases with donor size from 10 to 50% of the total intracellular label, and the median strand length of this fraction is half that of the donor strands. By combining these results with earlier data on the size dependence of the yield of transformants per unit of total intracellular donor label, we have calculated the probability that a marker in the intact fraction will be integrated, as a function of the length of the donor strand after entry. This probability has a linear dependence on strand length for activities below 40% of maximum, and extrapolates to zero activity at 77,000 daltons per strand.
将作为供体脱氧核糖核酸(DNA)引入肺炎双球菌感受态细胞的标记物的命运,在25℃进入细胞后立即根据供体DNA的大小进行了测定。发现部分标记物可溶于酸,部分已整合到染色体DNA中,显然是通过降解的供体DNA的重新整合,还有部分存在于长度小于输入供体DNA链的单链中。最后一部分显然构成了完整供体遗传标记整合的前体,被称为完整部分。对于大的供体DNA,完整部分包含细胞内总标记物的80%以上,但链长中位数已降至2.2×10⁶道尔顿。对于小的供体分子(每条链1×10⁵至6×10⁵道尔顿),完整部分随供体大小从细胞内总标记物的10%增加到50%,且该部分的链长中位数是供体链的一半。通过将这些结果与早期关于每单位细胞内总供体标记物转化子产量的大小依赖性数据相结合,我们计算了完整部分中的一个标记物在进入后作为供体链长度的函数被整合的概率。对于低于最大活性40%的活性,该概率与链长呈线性关系,并且在每条链77,000道尔顿处外推至零活性。