Ortuno M J, Lawther R P
Nucleic Acids Res. 1987 Feb 25;15(4):1521-42. doi: 10.1093/nar/15.4.1521.
Transcription in vitro of the regulatory region of the ilvGMEDA operon yields two attenuated RNAs initiated from the tandem promoters ilvGp1 and ilvGp2. Both S1 nuclease analysis and the fusion of ilvGp1 to galK indicate that transcription is not initiated in vivo from ilvGp1. However deletion of DNA sequences 150 to 100 bp upstream of ilvGp2 drastically reduces expression in vivo from ilvGp2. Both the distance separating ilvGp2 from the upstream DNA sequences and their relative orientation to each other on the DNA helix affect expression from ilvGp2. Deletion of DNA sequences approximately 400 bp upstream of ilvGp2 increases expression in vivo from this promoter. Analysis of products of transcription in vitro indicates that the effects observed in vivo are probably not due to DNA conformation or interactions of RNA polymerase.
ilvGMEDA操纵子调控区的体外转录产生了两种由串联启动子ilvGp1和ilvGp2起始的弱化RNA。S1核酸酶分析以及ilvGp1与galK的融合都表明,体内转录并非从ilvGp1起始。然而,ilvGp2上游150至100 bp的DNA序列缺失会大幅降低ilvGp2在体内的表达。ilvGp2与上游DNA序列之间的距离以及它们在DNA螺旋上彼此的相对方向都会影响ilvGp2的表达。ilvGp2上游约400 bp的DNA序列缺失会增加该启动子在体内的表达。体外转录产物分析表明,体内观察到的效应可能不是由于DNA构象或RNA聚合酶的相互作用。