Increasing DNA substrate specificity of the EcoDam DNA-(adenine N6)-methyltransferase by site-directed mutagenesis

被引：0
|
作者：
H. Elsawy
S. Chahar
机构：
[1] Tanta University,Chemistry Department, Faculty of Science
[2] King Faisal University,College of Science
[3] Al-Ahsaa,Department of Genetics, Rutgers
[4] The State University of New Jersey,undefined
来源：
Biochemistry (Moscow) | 2014年 / 79卷
关键词：
EcoDam; rational design; protein engineering;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
DNA methylation catalyzed by DNA methyltransferases (MTases) is widespread in prokaryotes. In an attempt to find EcoDam variants with enhanced preference for hemimethylated DNA, the L122, P134, and V133 residues were replaced with other amino acids using site directed mutagenesis, and the catalytic activity of all variants on unmethylated and hemimethylated substrates was studied. Our results showed that, in addition to L122A, the L122S and L122A/V133L EcoDam variants were able to sense the methylation status of the 5′-GATC-3′ double-stranded target recognition site and methylated only hemimethylated DNA.
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页码：1262 / 1266
页数：4
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