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PROCESS FOR COMMON CLONING GENES OF SAU961 RESTRICTIVE ENDONUCLEASE AND M.SAU961 MODIFYING METHYLASE AS WELL AS PROCESS FOR PURIFYING SAU961 RESTRICTIVE ENDONUCLEASE
PROCESS FOR COMMON CLONING GENES OF SAU961 RESTRICTIVE ENDONUCLEASE AND M.SAU961 MODIFYING METHYLASE AS WELL AS PROCESS FOR PURIFYING SAU961 RESTRICTIVE ENDONUCLEASE
Simultaneous cloning of genes of San 961 restrictive endonuclease and M. San 961 modificatory methylase enzymes and purification of Sau 961 restrictive endonuclease is given as: It is characteristic of cloning that DNS pref. 7596 DNS, is isolated from Staphylococcus aureus. This DNS is digested and coupled with a digested plasma-vector, pref. digested pBR 322 vector. The resultant DNS is ligated and introduced into Escherichia coli cells with transformation. The recombinant plasmid is re-septd. from the transformed cells. Isolated plasmid-DNS is subjected to exhaustive digestion with San 961 restrictive endonuclease. Escherichia coli cells are transfromed with digested plasmid DNS. These cells are extended on an agar plate contg. ampicillin. Finally plasmids are extracted from clones resistant to ampicillin. - For purificn. of San 961 restrictive endonuclease, clones of Escherichia coli, which carry genese of San 961 endonuclease and methylase are cultured in an ampicillin contg. medium. These cells are extracted and the suspn. of cells is disintegrated for centrifuging. IM sodium-chloride is added to the centrifuged supernatant, prior to gel-filtration. Fractions showing activity are combined and dialysed. San 961 endonuclease is isolated from the dialysed mix of adsorption, followed by elution.
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