Isolation and computer-aided characterization ofMmel, a Type II restriction endonuclease fromMethylophilus methylotrophus
- 1 January 1986
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 14 (13) , 5255-5274
- https://doi.org/10.1093/nar/14.13.5255
Abstract
A Type II restriction endonuclease, MmeI, has been purified from the obligate methylotroph, Methylophilus methylotrophus. The enzyme was shown to have the non-palindromic recognition sequence 5'-T C C Pu A C (N)20-3', 3'-A G G Py T G (N)18-5' and to cleave (as indicated) on the 3' side, generating a two nucleotide 3' projection. Determination of the recognition sequence was achieved using two new computer programs; RECOG, which predicts recognition sequences from the pattern of restriction fragments obtained from DNAs of known sequence, and GELSIM, which generates graphical simulations of DNA band patterns obtained by gel electrophoresis of restriction digests of sequenced DNA molecules.Keywords
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