Cloning of the Dam methyltransferase gene from Haemophilus influenzae bacteriophage HP1.

Acta microbiologica Polonica Pub Date : 1999-01-01
A Piekarowicz, J Bujnicki
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引用次数: 0

Abstract

The putative product of orf13 from the genome of Haemophilus influenzae HP1 bacteriophage shows homology only to bacteriophage T1 Dam methyltransferase, and a weak similarity to the conserved amino acids sequence motifs characteristic of m6A-methyltransferases. Especially interesting is lack of characteristic motif I responsible for binding of S-adenosylmethionine. Despite this fact, a DNA sequence of HP1 bacteriophage of Haemophilus influenzae encoding methyltransferase activity was cloned and expressed in Escherichia coli using pMPMT4 omega expression vector. The cloned methyltransferase recognizes the sequence 5'-GATC-3' and methylates an adenine residue. The enzyme methylates both double- and single-stranded DNA substrates.

流感嗜血杆菌噬菌体HP1 Dam甲基转移酶基因的克隆。
来自流感嗜血杆菌HP1噬菌体基因组的推测产物orf13仅与噬菌体T1 Dam甲基转移酶具有同源性,与m6a甲基转移酶的保守氨基酸序列基序特征具有弱相似性。特别有趣的是缺乏负责s -腺苷蛋氨酸结合的特征基序I。尽管如此,我们还是克隆了一段编码甲基转移酶活性的流感嗜血杆菌HP1噬菌体DNA序列,并利用pMPMT4 omega表达载体在大肠杆菌中表达。克隆的甲基转移酶识别序列5'- gtac -3'并甲基化腺嘌呤残基。这种酶可以甲基化双链和单链DNA底物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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