March 2019

cloning of GlaI site-specific methyl-directed DNA endonuclease gene and the comparison of the substrate specificity of recombinant and native enzymes is described. The analysis of recombinant GlaI endonuclease properties showed that the substrate specificity of native and recombinant enzymes didn't differ, but the concentration of recombinant enzyme exceeded 10 times of native enzyme

We have discovered a bacterial strain Lysinibacillus manganicus An22 that produces the new prototype of restriction endonuclease named LmnI. This enzyme recognizes a nonpalindromic DNA sequence 5’-GCTCC-3’/3’-CGAGG-5’. The LmnI restriction endonuclease preparation with the concentration of 1000 units/ml was isolated using four chromatographic steps. It was shown that new enzyme cuts its recognition sequence forming 3’-protruding ends as indicated by the arrows: 5’-GCTCCN↓-3’/3’-CGAG↑GN-5’, and can be applied to a IIS type of restriction-modification systems.