A Fluorometric Assay for DNA Cleavage Reactions Characterized with BamHI Restriction Endonuclease

1994 ◽  
Vol 220 (2) ◽  
pp. 377-383 ◽  
Author(s):  
S.P. Lee ◽  
D. Porter ◽  
J.G. Chirikjian ◽  
J.R. Knutson ◽  
M.K. Han
Keyword(s):  
Restriction Endonuclease ◽  
Dna Cleavage ◽  
Fluorometric Assay ◽  
Cleavage Reactions

A New Method for Determining the Stereochemistry of DNA Cleavage Reactions:  Application to theSfiI andHpaII Restriction Endonucleases and to the MuA Transposase†

Biochemistry ◽  
1999 ◽  
Vol 38 (14) ◽  
pp. 4640-4648 ◽  
Author(s):  
Kiyoshi Mizuuchi ◽  
Timothy J. Nobbs ◽  
Stephen E. Halford ◽  
Kenji Adzuma ◽  
Jun Qin
Keyword(s):  
Dna Cleavage ◽  
Restriction Endonucleases ◽  
New Method ◽  
Cleavage Reactions

DNA cleavage by the EcoRV restriction endonuclease: roles of divalent metal ions in specificity and catalysis

1999 ◽  
Vol 288 (1) ◽  
pp. 87-103 ◽  
Author(s):  
Geoffrey S. Baldwin ◽  
Richard B. Sessions ◽  
Symon G. Erskine ◽  
Stephen E. Halford
Keyword(s):  
Metal Ions ◽  
Restriction Endonuclease ◽  
Dna Cleavage ◽  
Divalent Metal ◽  
Divalent Metal Ions

scholarly journals Purification, properties and specificity of the restriction endonuclease from Bacillus stearothermophilus

1979 ◽  
Vol 177 (1) ◽  
pp. 49-62 ◽  
Author(s):  
C M Clarke ◽  
B S Hartley
Keyword(s):  
Enzyme Activity ◽  
Restriction Endonuclease ◽  
Dna Cleavage ◽  
Gel Electrophoresis ◽  
Bacillus Stearothermophilus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Major Protein ◽  
Activity Restriction

The restriction endonuclease BstI was purified from 70kg of Bacillus stearothermophilus. The final product is at least 97% pure as judged by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis; this major protein species co-migrates with the enzyme activity on native polyacrylamide-gel electrophoresis and isoelectric focusing. Pure restriction endonuclease BstI has a subunit mol.wt. of 26,000 and is probably a loosely associated dimer. The enzyme shows maximum activity at pH values between 7 and 9.5, and in the presence of 0.5-2mM-Mg2+. NaCl inhibits the restriction enzyme activity. Restriction endonuclease BstI cleaves DNA in a position identical with that cleaved by endonuclease BamHI (for Bacillus amyloliquefaciens), i.e.: (formula: see text). In the presence of high concentrations of enzyme, DNA cleavage occurs at secondary sites. This side-specificity is enhanced by the addition of glycerol. Preliminary studies indicate that these sites are of the type: (formula: see text).

Fingerprinting bacterial chromosomal DNA with restriction endonuclease EcoRI: comparison of Rhizobium spp. and identification of mutants

1979 ◽  
Vol 25 (7) ◽  
pp. 803-807 ◽  
Author(s):  
Jonathan R. Mielenz ◽  
L. E. Jackson ◽  
F. O'Gara ◽  
K. T. Shanmugam
Keyword(s):  
Restriction Endonuclease ◽  
Dna Cleavage ◽  
Rhizobium Trifolii ◽  
Chromosomal Dna ◽  
Mutant Strains ◽  
Restriction Endonuclease Ecori ◽  
Endonuclease Ecori ◽  
Cellular Dna ◽  
Derivatives Of ◽  
Japonicum Strain

Total cellular DNA from Rhizobium trifolii, R. meliloti, and R. japonicum strains 110 and 117 were prepared. DNA fragments generated with restriction endonuclease EcoRI from these DNA samples were compared in agarose gels after electrophoresis. DNA cleavage patterns generated from R. japonicum strain 110, R. trifolii, and R. meliloti were clearly distinguishable from each other. Restriction endonuclease cleavage patterns of DNA from R. japonicum strain 110 and presumptive R. trifolii mutant strains that nodulate soybean were found to be similar. Rhizobium trifolii mutant strains were also lysed by a phage specific for R. japonicum strain 110. These results show that "R. trifolii mutant strains" are indeed derivatives of R. japonicum strain 110 and not R. trifolii.

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