Site-specific endonucleases LplI and AagI
N. N. Sokolov, Z. P. Manalene, V. V. Butkus, A. B. Fitzner, E. B. Khoroshutina, A. A. Kalugin, A. A. Janulaitis
Institute of Biological and Medical Chemistry, Academy of Medical Sciences of the USSR, Moscow;All-Union Institute of Applied Enzymology, Vilnius
Abstract: New site-specific endonucleases LplI and AagI have been isolated from the Lacto-bacillus plantarum and Achromobacter agile cells, respectively. The enzymes' purification stages included treatment of cell-free extracts with polyethylenimine, fractionation in two-phase system by Albertsson's method, chromatography on blue Sepharose and DEAE-cellulose. The results of cleavage of a 5'-32P-labelled oligodeoxynucleotide duplex by restriction endonucleases LplI and AagI indicate that these enzymes recognize and cut the sequence AT↓CGAT, being therefore true isoschizomers of the ClaI restriction en-donuclease from Caryophanon latum. The L. plantarum strain has 400 fold endonuclease productivity as compared with the ClaI producent and is perspective for preparative-isolation, of LplI.
Russian Journal of Bioorganic Chemistry 1990, 16 (8):1040-1044