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淋病奈瑟菌中新DNA甲基转移酶编码基因的克隆与特性分析

Cloning and characterization of the gene encoding a new DNA methyltransferase from Neisseria gonorrhoeae.

作者信息

Radlińska M, Piekarowicz A

机构信息

Institute of Microbiology, University of Warsaw, Poland.

出版信息

Biol Chem. 1998 Nov;379(11):1391-5.

PMID:9865616
Abstract

A HindIII fragment of N. gonorrhoeae MS11 DNA coding for DNA methyltransferase (MTase) activity was cloned and expressed in E. coli AP1-200-9 cells. The sequence of 4681 bp was determined, and its analysis revealed two open reading frames (ORFs) sharing some similarity with known DNA MTases. ORF1 encodes an active N4mC MTase (M.NgoMV). The enzyme modifies only one strand of double stranded DNA and preferentially recognises the sequence GCCHR although it is able to methylate other sites. The exact recognition sequence cannot be precisely defined due to a relaxed specificity. The second ORF shows high homology to 5mC Mtases, but we were unable to demonstrate DNA methylating activity of its product either in vivo or in vitro.

摘要

编码DNA甲基转移酶(MTase)活性的淋病奈瑟菌MS11 DNA的HindIII片段被克隆并在大肠杆菌AP1-200-9细胞中表达。测定了4681 bp的序列,分析显示有两个开放阅读框(ORF)与已知的DNA MTase有一些相似性。ORF1编码一种活性N4mC MTase(M.NgoMV)。该酶仅修饰双链DNA的一条链,优先识别序列GCCHR,尽管它能够甲基化其他位点。由于特异性较宽松,无法精确确定确切的识别序列。第二个ORF与5mC Mtase具有高度同源性,但我们无法在体内或体外证明其产物的DNA甲基化活性。

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