منابع مشابه
Improved large-scale preparation of phage T4 endonuclease VII overexpressed in E. coli.
Using PCR, we cloned T4 gene 49, which encodes the endonuclease VII, and the inactive mutant gene 49 amE727 into vector pET-11a. In combination with Escherichia coli host strain BL21 (DE3), this system provided excellent repression of the expression of the highly toxic protein before induction with IPTG. After induction, the proteins were made in high quantities while remaining soluble. Dilutio...
متن کاملNear-simultaneous DNA cleavage by the subunits of the junction-resolving enzyme T4 endonuclease VII.
In common with a number of other DNA junction-resolving enzymes, endonuclease VII of bacteriophage T4 binds to a four-way DNA junction as a dimer, and cleaves two strands of the junction. We have used a supercoil-stabilized cruciform substrate to probe the simultaneity of cleavage at the two sites. Active endonuclease VII converts the supercoiled circular DNA directly into linear product, indic...
متن کاملScreening for mutations by enzyme mismatch cleavage with T4 endonuclease VII.
Each of four possible sets of mismatches (G.A/C.T, C.C/G.G, A.A/T.T, and C.A/G.T) containing the 8 possible single-base-pair mismatches derived from isolated mutations were examined to test the ability of T4 endonuclease VII to consistently detect mismatches in heteroduplexes. At least two examples of each set of mismatches were studied for cleavage in the complementary pairs of heteroduplexes ...
متن کاملAn apyrimidinic site kinks DNA and triggers incision by endonuclease VII of phage T4.
Apurinic/apyrimidinic lesions (AP-sites) occur frequently in DNA, generated by physically and chemically induced or spontaneous loss of bases. Repair mechanisms have evolved in organisms to deal efficiently with AP-sites by first incising the DNA at the lesion, followed by excision and resynthesis of the damaged strand. Here we report that endonuclease VII (endo VII) of phage T4, which was orig...
متن کاملT4 endonuclease involved in repair of DNA.
An enzyme activity, named T4 endonuclease V, was purified from T4-infected Escherichia coli. The enzyme induces single-stranded breaks in ultraviolet-irradiated DNA but does not act on native or heat-denatured DNA. The enzyme activity is dependent on the dose of ultraviolet irradiation, and the number of the breaks formed is approximately equal to the number of pyrimidine dimers present in the ...
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ژورنال
عنوان ژورنال: Journal of Biological Chemistry
سال: 1996
ISSN: 0021-9258
DOI: 10.1074/jbc.271.51.33148