Roger M
Proc Natl Acad Sci U S A. 1972 Feb;69(2):466-70. doi: 10.1073/pnas.69.2.466.
Heteroduplex molecules of pneumococcal DNA, prepared by cross-annealing resolved complementary strands, have been used as donors in transformation. A number of pairs of genetic markers are situated exclusively in the trans configuration in these heteroduplexes. Insertion of two markers from trans configuration into opposite DNA strands of a recipient genome should result in their segregation after one replication cycle. As a consequence, doubly transformed progeny would not appear, or would be markedly decreased. Contrary to these expectations, transformations with the heteroduplex DNAs give as many doubly transformed progeny for unlinked marker pairs as do homoduplex DNAs. For a pair of markers that normally are weakly linked, the frequencies of cotransfer are actually greater than those observed for a mixture of two singly marked homoduplex DNAs. These results lead to the conclusion that the heteroduplex DNAs are acted upon by repair enzymes of recipient cells so that markers introduced in the trans configuration are frequently converted to the cis configuration, either before or during integration. The efficiency of this conversion suggests that the repair and integration processes may be intimately connected. It is also concluded that complete breakdown of one strand of donor heteroduplex DNA does not occur during DNA uptake by recipient cells.
通过交叉退火分离出的互补链制备的肺炎球菌DNA异源双链分子已被用作转化的供体。在这些异源双链体中,许多对遗传标记仅以反式构型存在。将两个来自反式构型的标记插入受体基因组的相反DNA链中,在一个复制周期后应该会导致它们分离。因此,双转化后代不会出现,或者会显著减少。与这些预期相反,用异源双链DNA进行转化时,对于不连锁的标记对,产生的双转化后代与同源双链DNA一样多。对于通常弱连锁的一对标记,共转移频率实际上高于观察到的两种单标记同源双链DNA混合物的频率。这些结果得出结论,异源双链DNA受到受体细胞修复酶的作用,使得以反式构型引入的标记在整合之前或期间经常转化为顺式构型。这种转化效率表明修复和整合过程可能紧密相连。还得出结论,在受体细胞摄取DNA期间,供体异源双链DNA的一条链不会完全断裂。