Abstract
The genetic basis of the unique restriction endonuclease DpnI, that cleaves only at a methylated sequence, 5'-GmeATC-3', and of the complementary endonuclease DpnII, which cleaves at the same sequence when it is not methylated, was investigated. Different strains of Streptococcus pneumoniae isolated from patients contained either DpnI (two isolates) or DpnII (six isolates). The latter strains also contained DNA methylated at the 5'-GATC-3' sequence. A restrictable bacteriophage, HB-3, was used to characterize the various strains and to select for transformants. One laboratory strain contained neither DpnI nor Dpn II. It was probably derived from a DpnI-containing strain, and its DNA was not methylated at 5'-GATC-3'. Cells of this strain were transformed to the DpnI restriction phenotype by DNA from a DpnI-containing strain and to the DpnII restriction phenotype by DNA from a DpnII-containing strain. Neither cross-transformation, that is, transformation to one phenotype by DNA from a strain of the other phenotype, nor spontaneous conversion was observed. Extracts of transformants to the new restriction phenotype were shown to contain the corresponding endonuclease.
MeSH Terms
Bacteriophages/growth & development
DNA (Cytosine-5-)-Methyltransferases/metabolism
DNA Restriction Enzymes/genetics,metabolism
DNA, Bacterial/analysis
Deoxyribonucleases, Type II Site-Specific
Methylation
Streptococcus pneumoniae/enzymology,genetics
Transformation, Bacterial
Chemicals
DNA, Bacterial
DNA (Cytosine-5-)-Methyltransferases
DNA Restriction Enzymes
endodeoxyribonuclease DpnI
Deoxyribonucleases, Type II Site-Specific
GATC-specific type II deoxyribonucleases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Muckerman C C
Springhorn S S
Greenberg B
Lacks S A
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