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PMID: 14105 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Isolation and properties of a thermostable restriction endonuclease (ENDO R-Bst1503).

Journal of bacteriology ·Vol. 129 ·No. 2 ·1977-02-00 ·Pages 1110-20

Catterall JF, Welker NE

Abstract

A restriction endonuclease was isolated from Bacillus stearothermophilus1503-4R (Bst1503) and purified to homogeneity. The enzyme required Mg2+ ion as a cofactor. Bst1503 exhibited maximal activity between pH 7.5 and 8.0, between 60 and 65 degrees C, and with about 0.2 mM Mg2+. Bst1503 was not inactivated after exposure at 55 or 65 degrees C for up to 10 h. After 2 h of incubation at 70 degrees C, Bst1503 was inactivated by 65%. Bst1503 was rapidly inactivated at 75 degrees C. A single protein-staining band having a molecular weight of 46,000 was observed when Bst1503 was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was found to exist in two active forms, the predominating form with an S value of 8.3 (180,000) and the second form with an S value of 5.4 (96,000). No conversion between the 8.3S and 5.4S forms was observed after storage. Bst1503 recognized six sites in TP-1C deoxyribonucleic acid (DNA), one site in pSC101 and simian virus 40 DNAs, and three sites in lambdavir DNA. Bst1503 and BamHI were determined to be isoschizomers. The effect of temperatures on the activity and stability of BamHI was determined.

MeSH Terms
Bacteriophages DNA Restriction Enzymes/isolation & purification,metabolism DNA, Viral/metabolism Endonucleases/isolation & purification Geobacillus stearothermophilus/enzymology Hydrogen-Ion Concentration Magnesium/metabolism Molecular Weight Simian virus 40 Temperature
Chemicals
DNA, Viral Endonucleases DNA Restriction Enzymes Magnesium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Catterall J F
Welker N E
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39 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.129.2.1110-1120.1977
Published
1977-02-00
Pages
1110-20
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC235052
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.129.2.1110-1120.1977 PMC Full Text View journal details
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