Author:
Blumenthal R M,Gregory S A,Cooperider J S
Abstract
A 4.84-kilobase-pair plasmid was isolated from Proteus vulgaris (ATCC 13315) and cloned into the plasmid vector pBR322. Plasmid pBR322 contains substrate sites for the restriction endonucleases PvuI and PvuII. The recombinant plasmids were resistant to in vitro cleavage by PvuII but not PvuI endonuclease and were found to cause production of PvuII endonuclease or methylase activity or both in Escherichia coli HB101. The approximate endonuclease and methylase gene boundaries were determined through subcloning, Bal 31 resection, insertional inactivation, DNA-dependent translation, and partial DNA sequencing. The two genes are adjacent and appear to be divergently transcribed. Most E. coli strains tested were poorly transformed by the recombinant plasmids, and this was shown by subcloning and insertional inactivation to be due to the PvuII methylase gene. At a low frequency, stable methylase-producing transformants of a methylase-sensitive strain were obtained, and efficiently transformed cell mutants were isolated from them.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
Reference40 articles.
1. Linkage map of Escherichia coli K-12, edition 6;Bachmann B. J.;Microbiol. Rev.,1980
2. Variation in expression of sex factor genes in the Proteus-Providencia group relative to Escherichia coli;Baumberg S.;J. Bacteriol.,1975
3. Bickle T. A. 1982. The ATP-dependent restriction endonucleases p. 85-108. In S. M. Linn and R. J. Roberts (ed.) Nucleases. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
4. Plasmids of Escherichia coli as cloning vectors;Bolivar F.;Methods Enzymol.,1979
5. Construction and characterization of new cloning vehicles. I. A multipurpose system;Bolivar F.;Gene,1977
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