The Essential Carboxyl Group in Restriction Endonuclease EcoRI
نویسندگان
چکیده
منابع مشابه
Specificity of substrate recognition by the EcoRI restriction endonuclease.
The substrate specificity of the EcoRI restriction endonuclease can be varied in vitro by changing the pH and the ionic environment of the reaction. Phosphodiester bond cleavage occurs at a DNA hexanucleotide sequence d(N-G-A-A-T-T-C-N)/d(N-C-T-T-A-A-G-N) when the ionic strength is high, 100 mM Tris-HCl, 50 mM NaCl, 5 mM MgCl2, and the pH is approximately 7.3. Lowering the ionic strength to 25 ...
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The reactions of the EcoRI restriction endonuclease on the covalently closed DNA of plasmid pMB9 were studied in the presence of ethidium bromide. At the concentrations of ethidium bromide tested, which covered the range over which the DNA is changed from negatively to positively supercoiled, the dye caused no alteration to the rate at which this enzyme cleaved the covalently closed DNA to yiel...
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‘l’l~e SWYW EcoKI restriction endonuclease cleavage sites in bacteriophage P%d DNA have been mapped. The cleavage site map of circularly permut.ed P22 linear DNA is a circle. The positions of EcoRI sites in the early region of the P22 genomc were determined by comparing products of EcoRI digestion of maturtx liaear P22 chromosomes with the EcoRI cleavage fragments of DNA of three XimmP22 hybrid...
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Site-specific genetic recombinations promoted in vivo by the EcoRI endonuclease has been demonstrated by using constructed hybrid plasmids in which the chloramphenicol resistance gene was inactivated by insertion of DNA fragments at an EcoRI site within the gene. Such recombination can involve either the joining of intracellularly generated cohesive termini of the same DNA fragment or intermole...
متن کاملThe kinetic mechanism of EcoRI endonuclease.
Steady-state parameters governing cleavage of pBR322 DNA by EcoRI endonuclease are highly sensitive to ionic environment, with K(m) and k(cat) increasing 1,000-fold and 15-fold, respectively, when ionic strength is increased from 0.059 to 0.23 M. By contrast, pre-steady-state analysis has shown that recognition, as well as first and second strand cleavage events that occur once the enzyme has a...
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ژورنال
عنوان ژورنال: European Journal of Biochemistry
سال: 1981
ISSN: 0014-2956,1432-1033
DOI: 10.1111/j.1432-1033.1981.tb05678.x