Abstract
Overproduction of the NlaX DNA methyltransferase (M.NlaX) in an Escherichia coli host conferred resistance to SsoII restriction endonuclease (R.SsoII) digestion. This suggested an overlap of sequence specificity between M.NlaX and M.SsoII, the latter of which modifies the internal cytosine of the target sequence 5'-CCNGG-3'. A variant of M.NlaX (M.Sso/Nla), containing an N-terminal extension from M.SsoII, was also enzymatically active. Using deletion analysis, the N-terminal 71 amino-acid residues of M.SsoII were shown to be essential for modification activity.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Cloning, Molecular
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DNA Primers
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DNA-Cytosine Methylases / biosynthesis
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DNA-Cytosine Methylases / isolation & purification
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DNA-Cytosine Methylases / metabolism*
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Escherichia coli
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Kinetics
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Lactococcus lactis / enzymology*
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Lactococcus lactis / genetics
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Molecular Sequence Data
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Neisseria / enzymology*
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Neisseria / genetics
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Polymerase Chain Reaction
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Restriction Mapping
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Sequence Homology, Amino Acid
Substances
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DNA Primers
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Recombinant Proteins
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DNA modification methylase SsoII
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DNA-Cytosine Methylases