title: A counterselectable pACYC184-based lacZ alpha-complementing plasmid vector with novel multiple cloning sites; construction of chromosomal deletions in Klebsiella pneumoniae.
abstract: We have constructed a series of small, chloramphenicol-resistance-encoding, lacZ alpha-complementing vectors with novel multiple cloning sites, based on the pACYC184 replicon. The sacB gene of Bacillus subtilis, which is lethal to Gram- organisms in the presence of sucrose, was cloned into one of these, giving the counterselectable vector pSG335. This was used to substitute a streptomycin-resistance-encoding cassette for the ntrBC genes in the Klebsiella pneumoniae chromosome.
authors.affiliation: AFRC Institute of Plant Science Research, Nitrogen Fixation Laboratory, University of Sussex, Brighton, UK.
authors.forename: S
authors.initials: S
authors.lastname: Geissler
authors.forename: M
authors.initials: M
authors.lastname: Drummond
mesh_terms: D001483:Base Sequence; D002702:Chloramphenicol Resistance / Q000235:genetics; D002872:Chromosome Deletion; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D006602:Hexosyltransferases / Q000235:genetics; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D009591:Nitrogenase / Q000378:metabolism; D010957:Plasmids
chemical_list: D004269:DNA, Bacterial; D009591:Nitrogenase; D006602:Hexosyltransferases; C021513:levansucrase