Timmons M S, Lieb M, Deonier R C
Genetics. 1986 Aug;113(4):797-810. doi: 10.1093/genetics/113.4.797.
Intermolecular recombination between two IS5 elements was measured, using bacteriophage lambda recombination vectors, and was compared to recombination between two copies of an SV40 segment cloned into the same vectors. Experiments were conducted in the presence and in the absence of RecA and Red functions, and with the recombining inserts in the same or in reversed orientation. Under all conditions, IS5 elements recombined in a manner similar to the SV40 inserts, indicating that IS-encoded functions did not confer measurable additional intermolecular recombination ability to IS5 in E. coli K-12. Bacteriophages containing reversed IS5 inserts, for which the 16 base pair (bp) termini are identical in 15 positions and which display 12 bp of uninterrupted homology, recombined at approximately the same low frequency under Rec+ and Rec- conditions, indicating that these short homologies were not good substrates for the Rec system. Bacteriophages having reversed inserts recombined better under Red+ than under Red- conditions, but the crossovers were located in nonhomologous regions flanking the element termini. This suggests that 12-bp homologies are not good substrates for the Red system.
利用噬菌体λ重组载体测定了两个IS5元件之间的分子间重组,并将其与克隆到相同载体中的SV40片段的两个拷贝之间的重组进行了比较。实验在有和没有RecA和Red功能的情况下进行,并且重组插入片段处于相同或相反方向。在所有条件下,IS5元件的重组方式与SV40插入片段相似,这表明IS编码的功能并未赋予IS5在大肠杆菌K-12中可测量的额外分子间重组能力。含有反向IS5插入片段的噬菌体,其16个碱基对(bp)末端在15个位置相同,并且显示出12 bp的不间断同源性,在Rec+和Rec-条件下以大致相同的低频重组,这表明这些短同源性不是Rec系统的良好底物。具有反向插入片段的噬菌体在Red+条件下比在Red-条件下重组得更好,但交叉发生在元件末端侧翼的非同源区域。这表明12 bp的同源性不是Red系统的良好底物。