Weston-Hafer K, Berg D E
Department of Microbiology, Washington University Medical School, St. Louis, Missouri 63110.
Genetics. 1989 Apr;121(4):651-8. doi: 10.1093/genetics/121.4.651.
The contributions of direct and inverted repeats to deletion formation were studied by characterizing Ampr revertants of plasmids with a series of insertion mutations at a specific site in the pBR322 ampicillin resistance (amp) gene. The inserts at this site are palindromic, variable in length, and bracketed by 9- or 10-bp direct repeats of amp sequence. There is an additional direct repeat composed of 4 bp within the insert and 4 bp of adjoining amp sequence. DNA sequencing and colony hybridization of Ampr revertants showed that they contained either the parental amp sequence, implying deletion endpoints in the flanking 9- or 10-bp repeats, or a specific 1-bp substitution, implying endpoints in the 4-bp repeats. Although generally direct repeats seem to be used as deletion endpoints with a frequency proportional to their lengths, we found that with uninterrupted palindromes longer than 32 bp, the majority of deletions ended in the 4 bp, not the 9- or 10-bp repeats. This preferential use of the shorter direct repeats associated with palindromes is interpreted according to a DNA synthesis-error model in which hairpin structures formed by intrastrand pairing foster the slippage of nascent strands during DNA synthesis.
通过对pBR322氨苄青霉素抗性(amp)基因特定位点带有一系列插入突变的质粒的氨苄青霉素抗性(Ampr)回复子进行表征,研究了正向重复序列和反向重复序列对缺失形成的作用。该位点的插入序列为回文结构,长度可变,并由amp序列的9或10个碱基对的正向重复序列包围。插入序列内还有一个由4个碱基对组成的额外正向重复序列以及相邻amp序列的4个碱基对。对Ampr回复子的DNA测序和菌落杂交表明,它们要么包含亲本amp序列,这意味着缺失端点位于侧翼的9或10个碱基对的重复序列中,要么包含一个特定的1个碱基对的替换,这意味着缺失端点位于4个碱基对的重复序列中。尽管一般来说,正向重复序列似乎被用作缺失端点,其频率与它们的长度成正比,但我们发现,对于长度超过32个碱基对的不间断回文序列,大多数缺失的端点位于4个碱基对的重复序列中,而不是9或10个碱基对的重复序列中。根据DNA合成错误模型对与回文序列相关的较短正向重复序列的这种优先使用进行了解释,在该模型中,由链内配对形成的发夹结构促进了DNA合成过程中新生链的滑动。