Berget P B, Poteete A R, Sauer R T
J Mol Biol. 1983 Mar 15;164(4):561-72. doi: 10.1016/0022-2836(83)90050-5.
The structural gene for Salmonella bacteriophage P22 tail protein, gene 9, is separated from the remainder of the P22 late operon genes by the immI region. Early transcription of immI gene ant, which is immediately promoter proximal to gene 9, occurs in the same direction as late gene transcription but does not enter gene 9 coding sequences (Susskind & Youderian, 1982). We have cloned gene 9 and surrounding sequences into pBR322 and subsequently positioned lac UV5 promoters at varying distances before the start of gene 9 by DNA manipulations in vitro. Using an in vitro phage assembly assay to measure in vivo expression of tail protein from these plasmids and in vitro transcription reactions to measure transcriptional template activity of DNA fragments isolated from these plasmids, we have identified a region of DNA between gene ant and gene 9 that behaves as a transcription termination signal. The DNA sequence of this region shows hyphenated dyad symmetry followed by a run of seven thymine residues on the coding strand. This sequence can be drawn in a potential stem-and-loop secondary structure similar to known rho-independent transcription termination signal sequences. We discuss the role of this transcriptional terminator sequence in gene 9 expression and the early to late transcriptional switch in the P22 infection cycle.
沙门氏菌噬菌体P22尾蛋白的结构基因(基因9)与P22晚期操纵子的其余基因被immI区域隔开。紧邻基因9的启动子近端的immI基因ant的早期转录与晚期基因转录方向相同,但不进入基因9的编码序列(萨斯金德和尤德里安,1982年)。我们已将基因9及其周围序列克隆到pBR322中,随后通过体外DNA操作在基因9起始位点之前不同距离处定位lac UV5启动子。利用体外噬菌体组装试验来测量这些质粒中尾蛋白的体内表达,并利用体外转录反应来测量从这些质粒中分离的DNA片段的转录模板活性,我们确定了基因ant和基因9之间的一个DNA区域,该区域表现为转录终止信号。该区域的DNA序列显示出间断的二元对称性,随后在编码链上有七个胸腺嘧啶残基的连续排列。该序列可以形成类似于已知的不依赖ρ因子的转录终止信号序列的潜在茎环二级结构。我们讨论了这个转录终止序列在基因9表达以及P22感染周期中早期到晚期转录转换中的作用。