scholarly journals The SalGI restriction endonuclease. Mechanism of DNA cleavage

1982 ◽  
Vol 203 (1) ◽  
pp. 85-92 ◽  
Author(s):  
A Maxwell ◽  
S E Halford
Keyword(s):  
Restriction Endonuclease ◽  
Linear Form ◽  
Dna Cleavage ◽  
Tertiary Structure ◽  
Recognition Site ◽  
Optimal Conditions ◽  
Supercoiled Dna ◽  
Linear Forms ◽  
Open Circle ◽  
Concerted Reaction

The cleavage of supercoiled DNA of plasmid pMB9 by restriction endonuclease SalGI has been studied. Under the optimal conditions for this reaction, the only product is the linear form of the DNA, in which both strands of the duplex have been cleaved at the SalGI recognition site. DNA molecules cleaved in one strand at this site were found to be poor substrates for the SalGI enzyme. Thus, both strands of the DNA appear to be cleaved in a concerted reaction. However, under other conditions, the enzyme cleaves either one or both strands of the DNA; the supercoiled substrate is then converted to either open-circle or linear forms, the two being produced simultaneously rather than consecutively. We propose a mechanism for the SalGI restriction endonuclease which accounts for the reactions of this enzyme under both optimal and other conditions. These reactions were unaffected by the tertiary structure of the DNA.

scholarly journals The SalGI restriction endonuclease. Enzyme specificity

1982 ◽  
Vol 203 (1) ◽  
pp. 93-98 ◽  
Author(s):  
A Maxwell ◽  
S E Halford
Keyword(s):  
Restriction Endonuclease ◽  
Dna Sequences ◽  
Dna Cleavage ◽  
Competitive Inhibition ◽  
Recognition Site ◽  
Enzyme Specificity ◽  
Recognition Sequence ◽  
Kinetics Of ◽  
Endonuclease Enzyme

We have analysed the kinetics of DNA cleavage in the reaction between the SalGI restriction endonuclease and plasmid pMB9. This reaction is subject to competitive inhibition by DNA sequences outside the SalGI recognition site; we have determined the Km and Vmax. for the reaction of this enzyme at its recognition site and the KI for its interaction at other DNA sequences. We conclude that the specificity of DNA cleavage by the enzyme is only partly determined by the discrimination it shows for binding at its recognition sequence compared with binding to other DNA sequences.

scholarly journals On a characterization theorem on a-adic solenoids

2019 ◽  
Vol 489 (3) ◽  
pp. 227-231
Author(s):  
G. M. Feldman
Keyword(s):  
Gaussian Distribution ◽  
Real Line ◽  
Linear Form ◽  
Conditional Distribution ◽  
Random Variables ◽  
Characterization Theorem ◽  
The Other ◽  
Independent Random Variables ◽  
Linear Forms ◽  
The Real

According to the Heyde theorem the Gaussian distribution on the real line is characterized by the symmetry of the conditional distribution of one linear form of independent random variables given the other. We prove an analogue of this theorem for linear forms of two independent random variables taking values in an -adic solenoid containing no elements of order 2. Coefficients of the linear forms are topological automorphisms of the -adic solenoid.

Restriction endonuclease PshAI from Plesiomonas shigelloides with the novel recognition site 5′-GACNN/NNGTC

Gene ◽  
1990 ◽  
Vol 87 (1) ◽  
pp. 119-122 ◽  
Author(s):  
Michiko Miyahara ◽  
Katsuhisa Nakajima ◽  
Toshio Shimada ◽  
Katsutoshi Mise
Keyword(s):  
Restriction Endonuclease ◽  
Recognition Site ◽  
The Novel ◽  
Plesiomonas Shigelloides

scholarly journals Sau-PCR, a Novel Amplification Technique for Genetic Fingerprinting of Microorganisms

2005 ◽  
Vol 71 (10) ◽  
pp. 6401-6406 ◽  
Author(s):  
Viviana Corich ◽  
Alessandro Mattiazzi ◽  
Elisa Soldati ◽  
Angela Carraro ◽  
Alessio Giacomini
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Restriction Endonuclease ◽  
Genomic Dna ◽  
Recognition Site ◽  
Genetic Fingerprinting ◽  
Core Sequence ◽  
Amplification Technique

ABSTRACT The proposed technique is based on the digestion of genomic DNA with the restriction endonuclease Sau3AI and subsequent amplification with primers whose core sequence is based on the Sau3AI recognition site. The method was tested on strains of lactic acid bacteria but could be proposed for virtually any culturable organism from which DNA can be extracted.

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 The reactions of the EcoRi and other restriction endonucleases

1979 ◽  
Vol 179 (2) ◽  
pp. 353-365 ◽  
Author(s):  
S E Halford ◽  
N P Johnson ◽  
J Grinsted
Keyword(s):  
Restriction Endonuclease ◽  
Dna Sequences ◽  
Recognition Site ◽  
Single Strand ◽  
Restriction Endonucleases ◽  
Dna Duplex ◽  
Phosphodiester Bond ◽  
Linear Dna ◽  
Ecori Restriction ◽  
Dna Substrates

The reaction of the EcoRI restriction endonuclease was studied with both the plasmid pMB9 and DNA from bacteriophage lambda as the substrates. With both circular and linear DNA molecules, the only reaction catalysed by the EcoRI restriction endonuclease was the hydrolysis of the phosphodiester bond within one strand of the recognition site on the DNA duplex. The cleavage of both strands of the duplex was achieved only after two independent reactions, each involving a single-strand scission. The reactivity of the enzyme for single-strand scissions was the same for both the first and the second cleavage within its recognition site. No differences were observed between the mechanism of action on supercoiled and linear DNA substrates. Other restriction endonucleases were tested against plasmid pMB9. The HindIII restriction endonuclease cleaved DNA in the same manner as the EcoRI enzyme. However, in contrast with EcoRI, the Sa/I and the BamHI restriction endonucleases appeared to cleave both strands of the DNA duplex almost simultaneously. The function of symmetrical DNA sequences and the conformation of the DNA involved in these DNA–protein interactions are discussed in the light of these observations. The fact that the same reactions were observed on both supercoiled and linear DNA substrates implies that these interactions do not involve the unwinding of the duplex before catalysis.

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).

scholarly journals Characterization of a centromere-linked recombination hot spot in Saccharomyces cerevisiae.

1987 ◽  
Vol 7 (11) ◽  
pp. 3871-3879 ◽  
Author(s):  
M Neitz ◽  
J Carbon
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Restriction Fragment ◽  
Linear Form ◽  
Dna Hybridization ◽  
Hot Spot ◽  
Genetic Exchange ◽  
Linear Forms ◽  
Dna Fragment ◽  
Stimulation Of

A 1.5-kilobase-pair SalI-HindIII (SH) restriction fragment from the region of Saccharomyces cerevisiae chromosome XIV immediately adjacent to the centromere appears to contain sequences that act as a hot spot for mitotic recombination. The presence of SH DNA on an autonomously replicating plasmid stimulates homologous genetic exchange between yeast genomic sequences and those present on the plasmid. In all recombinants characterized, exchange occurs in plasmid yeast sequences adjacent to rather than within the SH DNA. Hybridization analyses reveal that SH-containing plasmids are present in linear as well as circular form in S. cerevisiae and that linear forms are generated by cleavage at specific sites. Presumably, it is the linear form of the plasmid that is responsible for the stimulation of genetic exchange. Based on these observations, it is proposed that this DNA fragment contains a centromere-linked recombination hot spot and that SH-stimulated recombination occurs via a mechanism similar to double-strand-gap repair (J. W. Szostak, T. Orr-Weaver, J. Rothstein, and F. Stahl, Cell 33:25-35 1983).

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