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噬菌体T7 DNA早期区域末端的转录终止位点。

The transcription termination site at the end of the early region of bacteriophage T7 DNA.

作者信息

Dunn J J, Studier F W

出版信息

Nucleic Acids Res. 1980 May 24;8(10):2119-32. doi: 10.1093/nar/8.10.2119.

Abstract

The nucleotide sequence through the transcription termination site for Escherichia coli RNA polymerase at the end of the early region of T7 DNA has been determined. RNA chains terminate at adjacent residues in the DNA sequence: about 2/3 of the chains end in C and 1/3 in G. A potential stem and loop structure, containing a stem of eight uninterrupted base pairs and a four-base loop, is centered 17-18 bases ahead of the chain termini. Transcription by E. coli RNA polymerase terminates efficiently at this site in vivo and in vitro, but transcription by T7 RNA polymerase is essentially unaffected. There are no primary cleavage sites for RNase III near the transcription termination site. The site of termination lies within HpaI fragment Q of T7 DNA, whose entire 446-nucleotide long sequence was determined. Cleavage sites for other restriction endonucleases are located conveniently for manipulating the DNA sequence around the termination site. The coding sequence for the last 82 amino acids of the T7 DNA ligase protein was identified, as was the beginning of a coding sequence for a possible late T7 protein from beyond the termination site.

摘要

已确定了T7 DNA早期区域末端大肠杆菌RNA聚合酶转录终止位点的核苷酸序列。RNA链在DNA序列中的相邻残基处终止:约2/3的链在C处终止,1/3在G处终止。一个潜在的茎环结构,包含一个由八个不间断碱基对组成的茎和一个四碱基环,位于链末端前方17 - 18个碱基处。大肠杆菌RNA聚合酶的转录在体内和体外均能在该位点有效终止,但T7 RNA聚合酶的转录基本不受影响。转录终止位点附近没有RNase III的主要切割位点。终止位点位于T7 DNA的HpaI片段Q内,其整个446个核苷酸长的序列已确定。其他限制性内切酶的切割位点位置方便,可用于操作终止位点周围的DNA序列。确定了T7 DNA连接酶蛋白最后82个氨基酸的编码序列,以及终止位点之外可能的晚期T7蛋白编码序列的起始部分。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e71d/324067/6d1e927bfc4b/nar00427-0013-a.jpg

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