A new site-specific endonuclease, ScaI, from Streptomyces caespitosus.
نویسندگان
چکیده
A new site-specific endonuclease has been isolated from Streptomyces caespitosus and named ScaI. Based on analysis of sequences around the restriction sites in pBR322 and pBR325, the recognition sequence of ScaI endonuclease was deduced to be a new hexanucleotide 5'-AGTACT-3'. The cleavage site was determined by comparing the ScaI-cleaved product of a primer-extended M13mp18-SCA DNA, which contains an AGTACT sequence, with dideoxy chain terminator ladders of the same DNA. ScaI was found to cleave the recognition sequence between the internal T and A, leaving flush ends to the cleaved fragments.
منابع مشابه
Interaction of the restriction endonuclease ScaI with its substrates.
The kinetic constants of the site-specific endonuclease, ScaI, for various substrates were determined. We estimated Vmax and Km for octa-, deca-, dodeca-, and hexadecanucleotides and for plasmid pBR322 DNA. Vmax for these substrates were close, but Km were quite different (in decreasing order, octa- greater than deca-, dodeca-, hexadeca- greater than pBR322). The results were discussed with res...
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ورودعنوان ژورنال:
- The Biochemical journal
دوره 231 1 شماره
صفحات -
تاریخ انتشار 1985