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噬菌体P1限制与修饰基因的转录分析

Transcriptional analysis of the restriction and modification genes of bacteriophage P1.

作者信息

Sharrocks A D, Hornby D P

机构信息

Department of Molecular Biology, Krebs Institute, University of Sheffield, UK.

出版信息

Mol Microbiol. 1991 Mar;5(3):685-94. doi: 10.1111/j.1365-2958.1991.tb00740.x.

Abstract

Bacteriophage P1 res and mod genes encode the restriction and modification polypeptides of the Type III restriction enzyme EcoP1. Northern blot analysis using res- and mod-specific probes revealed the presence of two separate transcripts in strains harbouring the EcoP1 restriction and modification genes. Furthermore, by constructing a series of fusions with a promoter less lacZ gene, we show that both the res and mod genes are transcribed from separate promoters. A more detailed investigation of the mod promoter region revealed two promoters located some 70 and 140bp upstream from the translational start codon. In addition, another pair of promoters and a further separate promoter are located more than 500bp upstream from this start codon. Two short open reading frames are located between these distal and proximal promoter clusters. Transcription of the res gene is initiated from within the mod open reading frame from two adjacent promoters. In addition a functional promoter is located on the antisense strand close to the res promoter region. The relationship between the transcription units of the res and mod genes is discussed.

摘要

噬菌体P1的res和mod基因编码III型限制酶EcoP1的限制和修饰多肽。使用res和mod特异性探针进行的Northern印迹分析显示,在携带EcoP1限制和修饰基因的菌株中存在两种独立的转录本。此外,通过构建一系列与无启动子lacZ基因的融合体,我们表明res和mod基因均从独立的启动子转录。对mod启动子区域进行更详细的研究发现,在翻译起始密码子上游约70和140bp处有两个启动子。此外,另一对启动子和一个单独的启动子位于该起始密码子上游500bp以上。在这些远端和近端启动子簇之间有两个短的开放阅读框。res基因的转录从mod开放阅读框内的两个相邻启动子起始。此外,一个功能性启动子位于反义链上靠近res启动子区域的位置。文中讨论了res和mod基因转录单元之间的关系。

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