噬菌体29早期启动子的核苷酸序列与转录

Nucleotide sequence and transcription of a bacteriophage 29 early promoter.

作者信息

Dobinson K F, Spiegelman G B

出版信息

J Biol Chem. 1985 May 25;260(10):5950-5.

Abstract

We have studied the in vitro and in vivo transcription of a promoter for Bacillus subtilis RNA polymerase on bacteriophage phi 29 DNA. The promoter is identified as an early promoter as it is transcribed in vitro by uninfected B. subtilis sigma 55-containing RNA polymerase; is transcribed in vivo at both 7 min after infection and in the presence of chloramphenicol; and is transcribed right to left on the standard phi 29 map. The nucleotide sequence of the promoter and the initiation site for RNA synthesis are reported. We have also examined the kinetics of RNA synthesis initiation using a single round run-off transcription assay. The overall rate of initiation was found to be 1.6 X 10(6) M-1 s-1 while the rate of conversion of unstable to stable polymerase-promoter complexes was 0.049 s-1. These values are comparable to those for similar promoters for Escherichia coli RNA polymerase.

摘要

我们研究了枯草芽孢杆菌RNA聚合酶在噬菌体phi 29 DNA上的一个启动子的体外和体内转录情况。该启动子被鉴定为早期启动子，因为它在体外能被未感染的含枯草芽孢杆菌σ55的RNA聚合酶转录；在感染后7分钟以及氯霉素存在的情况下在体内也能被转录；并且在标准的phi 29图谱上是从右向左转录。报告了该启动子的核苷酸序列以及RNA合成的起始位点。我们还使用单轮径流转录测定法研究了RNA合成起始的动力学。发现起始的总体速率为1.6×10⁶ M⁻¹ s⁻¹，而不稳定的聚合酶 - 启动子复合物转化为稳定复合物的速率为0.049 s⁻¹。这些值与大肠杆菌RNA聚合酶类似启动子的值相当。