THE RESTRICTION-ENDONUCLEASE RFLFII, ISOLATED FROM RUMINOCOCCUS-FLAVEFACIENS FD-1, RECOGNIZES THE SEQUENCE 5'-AGTACT-3', AND IS INHIBITED BY SITE-SPECIFIC ADENINE METHYLATION
M. Morrison et al., THE RESTRICTION-ENDONUCLEASE RFLFII, ISOLATED FROM RUMINOCOCCUS-FLAVEFACIENS FD-1, RECOGNIZES THE SEQUENCE 5'-AGTACT-3', AND IS INHIBITED BY SITE-SPECIFIC ADENINE METHYLATION, FEMS microbiology letters, 122(1-2), 1994, pp. 181-185
Molecular studies of the rumen bacterium Ruminococcus flavefaciens are
constrained by the lack of stable gene transfer systems. We report he
re on the characterization of RflFII, a restriction endonuclease isola
ted from R. flavefaciens FD-1. The enzyme is an isoschizomer of ScaI,
and cleavage of the DNA is blunt-ended, between the internal TA dinucl
eotide sequence of 5'-AGTACT-3'. Chromosomal DNA preparations were use
d to demonstrate that adenine methylation of DNA within the sequence 5
'-GTAC-3' inhibits both RflFII and the restriction endonucleases RsaI
and ScaI. Chromosomal DNA from R. flavefaciens FD-1 is also host modif
ied to protect against cleavage by ScaI.