The restriction endonuclease RflFII, isolated from Ruminococcus flavefaciens FD-1, recognizes the sequence 5'-AGTACT-3', and is inhibited by site-specific adenine methylation.
Molecular studies of the rumen bacterium Ruminococcus flavefaciens are constrained by the lack of stable gene transfer systems. We report here on the characterization of RflFII, a restriction endonuclease isolated from R. flavefaciens FD-1. The enzyme is an isoschizomer of ScaI, and cleavage of the DNA is blunt-ended, between the internal TA dinucleotide sequence of 5'-AGTACT-3'. Chromosomal DNA preparations were used 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 modified to protect against cleavage by ScaI.
Department of Animal Sciences, University of Illinois, Urbana-Champaign 61801.
M
M
Morrison
R I
RI
Mackie
B A
BA
White
D000225:Adenine / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D004262:DNA Restriction Enzymes / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D006095:Gram-Positive Cocci / Q000201:enzymology*; D008745:Methylation; D012417:Rumen / Q000382:microbiology*
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes; D000225:Adenine