title: Use of in vivo complementation in Mycobacterium tuberculosis to identify a genomic fragment associated with virulence.
abstract: Novel molecular tools and genetic methods were developed to isolate genomic fragments of Mycobacterium tuberculosis that may be associated with virulence. We sought to restore virulence, a characteristic of M. tuberculosis that is correlated with growth rate in mouse spleen and lung tissue, to the avirulent strain H37Ra by complementation. A representative library of the virulent M. tuberculosis strain H37Rv was constructed and transformed into H37Ra. Enrichment for individual faster-growing recombinants was achieved by passage of pools of H37Ra transformants harboring the H37Rv library through mice. A molecular strategy was devised to isolate and clone the H37Rv genomic DNA fragment ivg, which conferred a more rapid in vivo growth rate to H37Ra.
authors.affiliation: Department of Microbiology and Immunology, Howard Hughes Medical Institute, Albert Einstein College of Medicine, Bronx, New York 10461.
authors.forename: L
authors.initials: L
authors.lastname: Pascopella
authors.forename: F M
authors.initials: FM
authors.lastname: Collins
authors.forename: J M
authors.initials: JM
authors.lastname: Martin
authors.forename: M H
authors.initials: MH
authors.lastname: Lee
authors.forename: G F
authors.initials: GF
authors.lastname: Hatfull
authors.forename: C K
authors.initials: CK
authors.lastname: Stover
authors.forename: B R
authors.initials: BR
authors.lastname: Bloom
authors.forename: W R
authors.initials: WR
authors.lastname: Jacobs
mesh_terms: D000818:Animals; D002874:Chromosome Mapping; D015723:Gene Library; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D051379:Mice; D008810:Mice, Inbred C57BL; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D011995:Recombination, Genetic; D014774:Virulence