流感嗜血杆菌噬菌体HP1 Dam甲基转移酶基因的克隆。

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

摘要

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

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.

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