阿维链霉菌中一种独特的甲基特异性限制系统的表征
Characterization of a unique methyl-specific restriction system in Streptomyces avermitilis.
作者信息
MacNeil D J
机构信息
Exploratory Microbiology and Genetics, Merck Sharp & Dohme Research Laboratories, Rahway, New Jersey 07065.
出版信息
J Bacteriol. 1988 Dec;170(12):5607-12. doi: 10.1128/jb.170.12.5607-5612.1988.
Abstract
Streptomyces avermitilis contains a unique restriction system that restricts plasmid DNA containing N6-methyladenine or 5-methylcytosine. Shuttle vectors isolated from Escherichia coli RR1 or plasmids isolated from modification-proficient Streptomyces spp. cannot be directly introduced into S. avermitilis. This restriction barrier can be overcome by first transferring plasmids into Streptomyces lividans or a modification-deficient E. coli strain and then into S. avermitilis. The transformation frequency was reduced greater than 1,000-fold when plasmid DNA was modified by dam or TaqI methylases to contain N6-methyladenine or by AluI, HhaI, HphI methylases to contain 5-methylcytosine. Methyl-specific restriction appears to be common in Streptomyces spp., since either N6-methyladenine-specific or 5-methylcytosine-specific restriction was observed in seven of nine strains tested.
摘要
阿维链霉菌含有一种独特的限制系统，该系统会限制含有N6 - 甲基腺嘌呤或5 - 甲基胞嘧啶的质粒DNA。从大肠杆菌RR1分离得到的穿梭载体或从修饰能力正常的链霉菌属分离得到的质粒不能直接导入阿维链霉菌。这种限制屏障可以通过先将质粒转入变铅青链霉菌或修饰缺陷型大肠杆菌菌株，然后再转入阿维链霉菌来克服。当质粒DNA用dam或TaqI甲基化酶修饰以含有N6 - 甲基腺嘌呤，或用AluI、HhaI、HphI甲基化酶修饰以含有5 - 甲基胞嘧啶时，转化频率降低了1000倍以上。甲基特异性限制在链霉菌属中似乎很常见，因为在所测试的9个菌株中的7个中观察到了N6 - 甲基腺嘌呤特异性或5 - 甲基胞嘧啶特异性限制。
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