Positive selection procedure for entrapment of insertion sequence elements in gram-negative bacteria.
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.
D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D003470:Culture Media; D004251:DNA Transposable Elements; D006090:Gram-Negative Bacteria / Q000235:genetics*; D006602:Hexosyltransferases / Q000235:genetics; D010957:Plasmids; D012231:Rhizobium / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D012641:Selection, Genetic; D013395:Sucrose / Q000378:metabolism
D003470:Culture Media; D004251:DNA Transposable Elements; D013395:Sucrose; D006602:Hexosyltransferases; C021513:levansucrase