Huisman O, Fox M S
Genetics. 1986 Mar;112(3):409-20. doi: 10.1093/genetics/112.3.409.
Primary products of bacteriophage lambda recombination that display heterozygosity as a consequence of the presence of regions of heteroduplex DNA are rare in standard lambda crosses. Phage manifesting heterozygosity at a given allele are evident when recombinants, emerging from a cross, are selected for an exchange in a neighboring interval. We show that the abundance of such heterozygotes can be increased 10- to 20-fold by selection on an E. coli indicator that is defective in methyl-directed mismatch repair (mutL). Thus, the activity of the methyl-directed mismatch repair system is, at least in part, responsible for the low frequency of detectably heterozygous phage emerging from a standard cross. In a mutL indicator, many primary products of recombination are replicated without the intervention of mismatch repair.--The products of a six-factor phage cross have been plated on a mutL indicator allowing visual detection of those phage products heterozygous for one of the allelic pairs, cI. By genetic analysis, we show that the heteroduplex regions of these primary products of recombination are on the average about 4 kb in length and can include as much as half of the lambda genome.
在标准的λ噬菌体杂交中,由于存在异源双链DNA区域而表现出杂合性的噬菌体λ重组初级产物很少见。当从杂交中产生的重组体在相邻区间进行交换时被选择,在给定等位基因处表现出杂合性的噬菌体就很明显了。我们表明,通过在甲基定向错配修复(mutL)有缺陷的大肠杆菌指示菌上进行选择,这种杂合子的丰度可以增加10到20倍。因此,甲基定向错配修复系统的活性至少部分地导致了从标准杂交中出现的可检测到的杂合噬菌体的低频出现。在mutL指示菌中,许多重组初级产物在没有错配修复干预的情况下进行复制。——一个六因子噬菌体杂交的产物已被接种在mutL指示菌上,从而可以直观地检测出那些在等位基因对之一cI上呈杂合状态的噬菌体产物。通过遗传分析,我们表明这些重组初级产物的异源双链区域平均长度约为4kb,并且可以包含多达λ基因组的一半。