A site-specific endodeoxyribonuclease from Streptomyces albus CMI 52766 sharing site-specificity with Providencia stuartii endonuclease PstI

• Published in: Nucleic acids research Vol. 4; no. 6; pp. 1989 - 1998
• Main Author: Chater, K F
• Format: Journal Article
• Language: English
• Published: England 01.06.1977
• Subjects: Bacteriophages - metabolism; DNA Restriction Enzymes - isolation & purification; DNA Restriction Enzymes - metabolism; DNA, Viral - metabolism; Endonucleases - metabolism; Magnesium - pharmacology; Plasmids; Providencia - enzymology; Providencia stuartii; Sodium Chloride - pharmacology; Species Specificity; Streptomyces - enzymology; Streptomyces albus
• ISSN: 0305-1048; 1362-4962

A class II site-specific endodeoxyribonuclease (SalPI) was identified in cell-free extracts of Streptomyces albus CMI 52766 after high speed centrifugation and fractionation through Bio Gel AO.5M. SalPI cleaves lambda DNA into at least 18 fragments. Five cleavage sites were located in the linear lambda map by the use of double and triple restriction enzyme digests involving EcoRI, HindIII, SalGI and another new Streptomyces enzyme, SacI. The results were indistinguishable from those previously obtained for a Providencia stuartii enzyme, PstI, by Smith, Blattner & Davies (Nucleic Acids Res. 1976 3, 343). SalPI and PstI were shown by a double digest test to have the same site specificity. None of 34 phages tested was obviously restricted by S. albus CMI 52766, and correspondingly DNA from two of them was not cleaved in vitro by SalPI. DNA from Streptomyces phage that does not form plaques on S. albus CMI 52766, and plasmid SCP2 DNA from S. coelicolor A3 (2), were both cleaved.