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PMID: 3029675 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Nucleotide sequence and expression of the gene encoding the EcoRII modification enzyme.

Nucleic acids research ·Vol. 15 ·No. 1 ·1987-01-12 ·Pages 313-32

Som S, Bhagwat AS, Friedman S

Abstract

The gene coding for the EcoRII modification enzyme has been cloned and the nucleotide sequence of 1933 base pairs containing the gene has been determined. The gene codes for a protein of 477 amino acids. Two transcriptional start sites have been mapped by S1 mapping. One deletion that removes 34 N-terminal amino acids was found to have partial enzyme activity. Comparison of the EcoRII methylase sequence with other cytosine methylases revealed several domains of partial homology among all cytosine methylases. Cloning the gene in multicopy pUC vectors increased the expression by 6-18 fold. A 40 fold overproduction of the EcoRII methylase was obtained by cloning the gene in the expression vector carrying the lambda PL promoter.

MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular DNA Restriction Enzymes/genetics,metabolism Deoxyribonucleases, Type II Site-Specific Escherichia coli/enzymology,genetics Genes Genes, Bacterial Genetic Vectors Plasmids Transcription, Genetic
Chemicals
DNA Restriction Enzymes CCWGG-specific type II deoxyribonucleases Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Som S
Bhagwat A S
Friedman S
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43 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/15.1.313
Published
1987-01-12
Pages
313-32
Language
English
Region
England
NLM ID
0411011
PMCID
PMC340412
Subset
IM
Grants
NIGMS NIH HHS · GM27787 · United States
Databases
GENBANK
X05050
External Links
PubMed source DOI: 10.1093/nar/15.1.313 PMC Full Text View journal details
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