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Molecular cloning and transposition of a kanamycin resistance determinant fromCampylobacter jejunibetween replicons inEscherichia coli

✍ Scribed by R. Son; A. Ansary

Publisher: Springer
Year: 1996
Tongue: English
Weight: 449 KB
Volume: 12
Category: Article
ISSN: 1573-0972
DOI: 10.1007/bf00340210

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✦ Synopsis

This paper reports a restriction map of a fragment of DNA encoding kanamycin resistance cloned from plasmid DNA of Campylobacter jejuni ABA94 in the recombinant plasmid pRS9421-1. In transposition experiments, kanamycin-resistant R751::km9421 transconjugants appeared at frequencies of 10(-7) per donor cell. These transconjugants harboured a plasmid 4 kb larger than the parental 49 kb plasmid R751. Restriction enzyme analysis and Southern blot hybridization of these transconjugants showed that the kanamycin resistant determinant had transposed from recombinant plasmid pRS9421-1 to plasmid R751.

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The erythromycin resistance gene (Em(r)) from Campylobacter jejuni ABA94 plasmid DNA was cloned into the pUC18 vector and then expressed in Escherichia coli. The location of the Em(r) determinant on the chimeric plasmid was determined by restriction endonuclease mapping within a 0.8-kb EcoRI fragmen