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J Bacteriol. Nov 1985; 164(2): 918–921.
PMCID: PMC214340
Positive selection procedure for entrapment of insertion sequence elements in gram-negative bacteria.
P Gay, D Le Coq, M Steinmetz, T Berkelman, and C I Kado
Abstract
We constructed the broad-host-range plasmid pUCD800 containing the sacB gene of Bacillus subtilis for use in the positive selection and isolation of insertion sequence (IS) elements in gram-negative bacteria. Cells containing pUCD800 do not grow on medium containing 5% sucrose unless the sacB gene is inactivated. By using pUCD800, we isolated a 1.4-kilobase putative IS element from Agrobacterium tumefaciens NT1RE by selection for growth on sucrose medium. This putative IS element appears to be unique to Agrobacterium strains.
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