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Gene ; 157(1-2): 43-7, 1995 May 19.
Article in English | MEDLINE | ID: mdl-7607523

ABSTRACT

We have characterized a family of related restriction-modification (R-M) systems from the soil bacterium Herpetosiphon giganteus (Hgi). A comparison of their genetic organization reveals two types of regulatory proteins, called controlling ORF C. While one of these small reading frames derived from RM.HgiCI seems to be an enhancer of its own promoter, evidence is provided for a silencer function of the other ORF C derived from the closely related AvaII-type systems RM.HgiBI/CII/EI. The respective silencer function is detected during our various attempts to clone three isoschizomers with unusually high differences in their specific activity. Sequencing and site-directed mutagenesis revealed just two amino acids as being responsible for a massive increase in specific activity of these endonucleases.


Subject(s)
DNA Restriction Enzymes/biosynthesis , Deoxyribonucleases, Type II Site-Specific/biosynthesis , Gene Expression , Genes, Bacterial , Base Sequence , Cloning, Molecular/methods , DNA Restriction Enzymes/genetics , DNA Restriction Enzymes/metabolism , Deoxyribonucleases, Type II Site-Specific/genetics , Deoxyribonucleases, Type II Site-Specific/metabolism , Gene Expression Regulation, Enzymologic , Molecular Sequence Data , Restriction Mapping , Substrate Specificity
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