title: Construction of a nontypeable Haemophilus influenzae-specific ectopic delivery vector.
abstract: Complementation of chromosomal mutations in trans can introduce artifacts due to the number of episomal copies of the gene in question. One solution is to study the gene expressed at a single ectopic site in cis. We have designed and constructed a vector that allows homologous recombination into a gene encoding a frame-shifted IS1016-V6 protein in the Haemophilus influenzae Rd KW20 chromosome (HI1018). This site is the location of the > or = 35 kilobase capsule locus in encapsulated type b and d strains. This locus is not present in the nontypeable Rd KW20 strain, thus allowing ectopic expression of genes homologously recombined into HI1018 without polar effects.
authors.affiliation: Seattle Biomedical Research Institute, 4 Nickerson St. Suite 200, Seattle, WA 98109, USA. daines2@llnl.gov
authors.forename: Dayle A
authors.initials: DA
authors.lastname: Daines
authors.forename: Arnold L
authors.initials: AL
authors.lastname: Smith
mesh_terms: D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D016368:Frameshift Mutation; D005798:Genes, Bacterial / Q000235:genetics*; D005822:Genetic Vectors / Q000737:chemistry*; D006193:Haemophilus influenzae / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic
chemical_list: D004269:DNA, Bacterial