Cloning the modification methylase gene of Bacillus sphaericus R in Escherichia coli |
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Authors: | E Szomolányi A Kiss P Venetianer |
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Institution: | Institute of Biochemistry, Biological Research Center of the Hungarian Academy of Sciences, H-6 701 Szeged P.O.B. 521 Hungary |
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Abstract: | The gene coding for the sequence-specific modification methylase methM . BspI of Bacillus sphaericus R has been cloned in Escherichia coli by means of plasmid pBR322. The selection was based on the expression of the cloned gene which rendered the recombinant plasmid resistant to BspI restriction endonuclease cleavage. The gene is carried by a 9 kb BamHI fragment and by a smaller 2.5 kb EcoRI fragment derived from the BamHI fragment. The Bsp-specific methylase level was found to be higher in the recombinant clones than in the parental strain. The methylase gene is probably located on the Bacillus sphaericus chromosome, and not on a plasmid known to be carried by this strain. The recombinant clones do not exhibit an BspI restriction endonuclease activity. |
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Keywords: | pBR322 plasmid expression in foreign host kb kilobase pairs Md megadaltons mol wt molecular weight(s) |
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