凝结芽孢杆菌K中位点特异性DNA甲基转移酶的分离与鉴定
Isolation and characterization of site-specific DNA-methyltransferases from Bacillus coagulans K.
作者信息
Svadbina I V, Zelinskaya N V, Kovalevskaya N P, Zheleznaya L A, Matvienko N I
机构信息
Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino 142290, Moscow Region, Russia.
出版信息
Biochemistry (Mosc). 2004 Mar;69(3):299-305. doi: 10.1023/b:biry.0000022061.29918.8b.
Two site-specific DNA methyltransferases, M.BcoKIA and M.BcoKIB, were isolated from the thermophilic strain Bacillus coagulans K. Each of the methylases protects the recognition site 5'-CTCTTC-3'/5'-GAAGAG-3' from cleavage with the cognate restriction endonuclease BcoKI. It is shown that M.BcoKIB is an N6-adenine specific methylase and M.BcoKIA is an N4-cytosine specific methylase. According to bisulfite mapping, M.BcoKIA methylates the first cytosine in the sequence 5'-CTCTTC-3'.
从嗜热菌株凝结芽孢杆菌K中分离出两种位点特异性DNA甲基转移酶，即M.BcoKIA和M.BcoKIB。每种甲基化酶都能保护识别位点5'-CTCTTC-3'/5'-GAAGAG-3'不被同源限制性内切酶BcoKI切割。结果表明，M.BcoKIB是一种N6-腺嘌呤特异性甲基化酶，而M.BcoKIA是一种N4-胞嘧啶特异性甲基化酶。根据亚硫酸氢盐图谱分析，M.BcoKIA使序列5'-CTCTTC-3'中的第一个胞嘧啶发生甲基化。
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