F质粒转移操纵子的控制区：traJ和traY基因的DNA序列以及traY至Z启动子的特性分析

The control region of the F plasmid transfer operon: DNA sequence of the traJ and traY genes and characterisation of the traY leads to Z promoter.

作者信息

Fowler T, Taylor L, Thompson R

出版信息

Gene. 1983 Dec;26(1):79-89. doi: 10.1016/0378-1119(83)90038-0.

Abstract

The complete nucleotide sequence of the F plasmid transfer genes traJ and traY, together with the promoter-proximal region of the traA gene has been determined. The traJ reading frame has been confirmed by sequencing the traJ90 amber mutant allele. The predicted amino acid sequence of the TraJ protein shows that this outer-membrane protein lacks a signal sequence. The pattern of codon usage within the traJ gene is different from that of genes for abundant outer-membrane proteins and is closer to that of genes that are expressed at relatively low levels. We have located the traY leads to Z operon promoter by in vitro run-off transcription experiments and have developed in vivo assays for the activity of the promoter by fusing it to galactokinase and kanamycin-resistance genes.

摘要

已确定F质粒转移基因traJ和traY的完整核苷酸序列，以及traA基因的启动子近端区域。通过对traJ90琥珀突变等位基因进行测序，证实了traJ阅读框。TraJ蛋白的预测氨基酸序列表明，这种外膜蛋白缺乏信号序列。traJ基因内的密码子使用模式不同于丰富外膜蛋白的基因，更接近于相对低水平表达的基因。我们通过体外 runoff转录实验定位了traY导致Z操纵子启动子，并通过将其与半乳糖激酶和卡那霉素抗性基因融合，开发了体内启动子活性测定方法。