Purification of BsuE methyltransferase and its application in genome mapping.
We have used a combination of BsuE methyltransferase (M-BsuE) and NotI restriction enzyme to cut genomic DNA at a subset of NotI sites. The usefulness of this system is shown in a re-examination of the restriction map of the human MHC. Combinations of methylases and restriction enzymes can be used to generate cuts at different frequencies in genomic DNA, such that they generate ends complementary to NotI ends, and can be used in conjunction with NotI linking clones in chromosome jumping experiments. These enzyme combinations have the potential to produce cutting sites in genomic DNA spaced at intervals favorable for extensive mapping, fragment enrichment, and cloning efforts.
Shukla
H
H
Yale University School of Medicine, Department of Human Genetics, New Haven, CT.
Kobayashi
Y
Y
Arenstorf
H
H
Yasukochi
Y
Y
Weissman
S M
SM
D001483:Base Sequence; D015139:Blotting, Southern; D002461:Cell Line, Transformed; D002845:Chromatography; D002874:Chromosome Mapping; D015257:DNA-Cytosine Methylases / Q000302:isolation & purification / Q000378:metabolism*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008285:Major Histocompatibility Complex / Q000235:genetics; D008969:Molecular Sequence Data; D015183:Restriction Mapping
C055352:DNA modification methylase BsuE; D015257:DNA-Cytosine Methylases; D015252:Deoxyribonucleases, Type II Site-Specific; C038565:GCGGCCGC-specific type II deoxyribonucleases