Ueno T, Ito H, Kimizuka F, Kotani H, Nakajima K
Bioproducts Development Center, Takara Shuzo Co., Ltd., Shiga, Japan.
Nucleic Acids Res. 1993 May 25;21(10):2309-13. doi: 10.1093/nar/21.10.2309.
The genes from Moraxella bovis encoding the MboI restriction--modification system were cloned and expressed in Escherichia coli. Three open reading frames were found in the sequence containing the genes. These genes, which we named mboA, mboB, and mboC, had the same orientation in the genome. Genes mboA and mboC encoded MboI methyltransferases (named M.MboA and M.MboC) with 294 and 273 amino acid residues, respectively. The mboB gene coded for MboI restriction endonuclease (R.MboI) with 280 amino acid residues. Recombinant E.coli-MBOI, which contained the whole MboI system, overproduced R.MboI. R.MboI activity from E.coli-MBOI was 480-fold that of M.bovis. The amino acid sequences deduced from these genes were compared with those of other restriction--modification systems. The protein sequences of the MboI system had 38-49% homology with those of the DpnII system.
编码莫拉克斯氏菌属牛源MboI限制 - 修饰系统的基因被克隆并在大肠杆菌中表达。在包含这些基因的序列中发现了三个开放阅读框。我们将这些基因命名为mboA、mboB和mboC，它们在基因组中具有相同的方向。基因mboA和mboC分别编码具有294和273个氨基酸残基的MboI甲基转移酶（分别命名为M.MboA和M.MboC）。mboB基因编码具有280个氨基酸残基的MboI限制性内切酶（R.MboI）。含有完整MboI系统的重组大肠杆菌 - MBOI过量产生R.MboI。来自大肠杆菌 - MBOI的R.MboI活性是牛莫拉克斯氏菌的480倍。将从这些基因推导的氨基酸序列与其他限制 - 修饰系统的序列进行比较。MboI系统的蛋白质序列与DpnII系统的蛋白质序列具有38 - 49％的同源性。
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