CLONING THE BAMHI RESTRICTION MODIFICATION SYSTEM
CLONING THE BAMHI RESTRICTION MODIFICATION SYSTEM
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DOI：
10.1093/nar/17.3.979
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发表时间：
1989-02-11
影响因子：
14.9
通讯作者：
NWANKWO, DO
中科院分区：
文献类型：
--
作者：
BROOKS, JE;BENNER, JS;NWANKWO, DO
BamHI，一种来自解淀粉芽孢杆菌H的II型限制性修饰系统，识别序列GGATCC。甲基化酶和核酸内切酶基因已经在单独的步骤中克隆到大肠杆菌中;该克隆能够限制未修饰的噬菌体。尽管在克隆中甲基化酶和内切核酸酶基因存在于相同的pACYC 184载体上，但该系统可以在E.大肠杆菌中的甲基化酶基因的额外拷贝存在于单独的载体上。对BamHI甲基化酶活性的初始选择也产生了第二个BamHI甲基化酶基因，该基因在DNA序列上不同源，并且与核酸内切酶连接的甲基化酶基因相比与不同的基因组限制性片段杂交。最后，研究了BamHI系统与E.大肠杆菌Dam和Mcr A和B的功能，已被研究，并在这里报告。
BamHI, a Type II restriction modification system from Bacillus amyloliquefaciensH recognizes the sequence GGATCC. The methylase and endonuclease genes have been cloned into Escherichia coli in separate steps; the clone is able to restrict unmodified phage. Although within the clone the methylase and endonuclease genes are present on the same pACYC184 vector, the system can be maintained in E. coli only with an additional copy of the methylase gene present on a separate vector. The initial selection for BamHI methylase activity also yielded a second BamHI methylase gene which is not homologous in DNA sequence and hydbridizes to different genomic restriction fragments than does the endonuclease-linked methylase gene. Finally, the interaction of the BamHI system with the E. coli Dam and the Mcr A and B functions, have been studied and are reported here.