Moyle H, Waldburger C, Susskind M M
Department of Biological Sciences, University of Southern California, Los Angeles 90089-1340.
J Bacteriol. 1991 Mar;173(6):1944-50. doi: 10.1128/jb.173.6.1944-1950.1991.
Oligonucleotide-directed mutagenesis was used to complete a collection of mutations in the -35 and -10 hexamers of the ant promoter of Salmonella phage P22. The effects of all 36 single-base-pair substitutions on promoter strength in vivo were measured in strains carrying the mutant promoters fused to an ant-lacZ gene on a single-copy prophage. The results of these assays show that certain consensus base pairs are more important than others; in general, the least-critical positions are among the most poorly conserved. Some mutations within the hexamers have smaller effects on promoter strength than certain mutations outside the hexamers in this and other promoters. Several different patterns of base pair preferences are observed. These hierarchies of base pair preferences correlate well (but not perfectly) with the hierarchies defined by the frequency distribution of base pairs at each position among wild-type promoters. The hierarchies observed in the ant promoter also agree well with most of the available information on base pair preferences in other promoters.
利用寡核苷酸定向诱变技术,完成了沙门氏菌噬菌体P22 ant启动子-35和-10六聚体中一系列突变。在携带与单拷贝原噬菌体上的ant-lacZ基因融合的突变启动子的菌株中,测定了所有36个单碱基对替换对体内启动子强度的影响。这些检测结果表明,某些共有碱基对比其他碱基对更重要;一般来说,最不重要的位置是保守性最差的位置。在该启动子以及其他启动子中,六聚体内的一些突变对启动子强度的影响小于六聚体外的某些突变。观察到几种不同的碱基对偏好模式。这些碱基对偏好层次与野生型启动子中每个位置的碱基对频率分布所定义的层次(但不完全)高度相关。在ant启动子中观察到的层次也与其他启动子中碱基对偏好的大多数现有信息高度一致。