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PMID: 2827113 Published · ppublish English Journal Article

Cloning, sequencing and expression of the Taq I restriction-modification system.

Nucleic acids research ·Vol. 15 ·No. 23 ·1987-12-10 ·Pages 9781-96

Slatko BE, Benner JS, Jager-Quinton T, Moran LS, Simcox TG, Van Cott EM, Wilson GG

Abstract

The Taq I modification and restriction genes (recognition sequence TCGA) have been cloned in E. coli and their DNA sequences have been determined. Both proteins were characterized and the N-terminal sequence of the endonuclease was determined. The genes have the same transcriptional orientation with the methylase gene 5' to the endonuclease gene. The methylase gene is 1089 bp in length (363 amino acids, 40,576 daltons); the endonuclease gene is 702 bp in length (234 amino acids, 27,523 daltons); they are separated by 132 bp. Both methylase and endonuclease activity can be detected in cell extracts. The clones fully modify the vector and chromosomal DNA but they fail to restrict infecting phage. Clones carrying only the restriction gene are viable even in the absence of modification. The restriction gene contains 7 Taq I sites; the modification gene contains none. This asymmetric distribution of sites could be important in the regulation of the expression of the endonuclease gene.

MeSH Terms
Amino Acid Sequence Bacteriophage lambda/genetics Base Sequence Chromosome Mapping Cloning, Molecular/methods Codon DNA Restriction Enzymes/genetics,isolation & purification DNA, Bacterial/genetics,isolation & purification Deoxyribonucleases, Type II Site-Specific Escherichia coli/enzymology,genetics Genes, Bacterial Molecular Sequence Data Species Specificity Thermus/enzymology,genetics
Chemicals
Codon DNA, Bacterial DNA Restriction Enzymes Deoxyribonucleases, Type II Site-Specific TCGA-specific type II deoxyribonucleases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Slatko B E
New England Biolabs, Inc., Beverly, MA 01915.
Benner J S
Jager-Quinton T
Moran L S
Simcox T G
Van Cott E M
Wilson G G
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/15.23.9781
Published
1987-12-10
Pages
9781-96
Language
English
Region
England
NLM ID
0411011
PMCID
PMC306531
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/15.23.9781 PMC Full Text View journal details
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