Effect of mercury substitution of DNA on its susceptibility to cleavage by restriction endonucleases.
Mercurated DNA was synthesized in vitro by substituting Hg-dCTP or Hg-dUMP for dCTP in one strand of M13mp8DNA in a DNA polymerase I reaction. Restriction enzymes, including Sma I, Pst I, Bam HI, Hind III, and Hinc II, were completely inactive when their recognition sites were fully substituted with Hg-dCMP, while Hg-dUMP containing DNA was hydroxlyzed to some extent. Under conditions favoring star activities, or when DNA was substituted with a low level of mercury-nucleotide, DNA was cleaved by restriction enzymes. Susceptibility to degrading and synthesizing enzymes and insensitivity to restriction endonucleases of fully mercurated DNA makes mercuration an attractive molecular "tag" for in vitro manipulation and selective isolation of Hg-DNA.
Banfalvi
G
G
Department I, Semmelweis University Medical School, Budapest, Hungary.
Sarkar
N
N
D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D004262:DNA Restriction Enzymes / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D008628:Mercury / Q000737:chemistry*; D008969:Molecular Sequence Data
D004247:DNA; D004262:DNA Restriction Enzymes; D008628:Mercury