Benson S A, DeCloux A M, Munro J
Department of Microbiology, University of Maryland, College Park 20742-4451.
Genetics. 1991 Nov;129(3):647-58. doi: 10.1093/genetics/129.3.647.
We have characterized a nonlethal selection for mutations that allow Escherichia coli to grow on large maltodextrins (Dex+) in the absence of the lamB encoded maltoporin LamB. These Dex+ mutations occur before and after imposition of the selection and the selection does not result in a general increase in mutagenesis. The recovered Dex+ mutations are almost exclusively mutations that alter the ompF gene that encodes a major E. coli porin, OmpF even though analogous mutations in the homologous ompC gene, which encodes the OmpC porin, can confer a Dex+ phenotype. We show that the bias for ompF mutations results from a biased recovery and that the genetic background of the starting strain and the selection itself influences the type of mutants that are recovered. When we use a strain carrying an amber mutation in the lamB gene we observe the same preference for ompF mutations as when we start with a lamB deletion strain. In addition, we show that there is no preferential mutagenesis of the lamB gene during the selection which induces transcription of the lamB gene. We present evidence that the biased recovery of mutants observed in this selection does not result from adaptive or directed mutagenesis and that the phenotypic fitness which allows recovery of Dex+ mutants involves more than the increased ability to take up maltodextrins.
我们已鉴定出一种针对突变的非致死性筛选方法,该方法可使大肠杆菌在缺乏由lamB编码的麦芽糖孔蛋白LamB的情况下,在大型麦芽糖糊精(Dex+)上生长。这些Dex+突变在施加筛选之前和之后都会发生,并且该筛选不会导致诱变作用普遍增加。回收的Dex+突变几乎都是改变编码大肠杆菌主要孔蛋白OmpF的ompF基因的突变,尽管编码OmpC孔蛋白的同源ompC基因中的类似突变也可赋予Dex+表型。我们表明,对ompF突变的偏好是由于回收偏差导致的,并且起始菌株的遗传背景和筛选本身会影响回收的突变体类型。当我们使用在lamB基因中携带琥珀突变的菌株时,我们观察到对ompF突变的偏好与从lamB缺失菌株开始时相同。此外,我们表明在诱导lamB基因转录的筛选过程中,lamB基因没有优先诱变。我们提供的证据表明,在此筛选中观察到的突变体回收偏差并非由适应性或定向诱变导致,并且允许回收Dex+突变体的表型适应性涉及的不仅仅是摄取麦芽糖糊精能力的提高。