title: Molecular characterization of heterologous HIV-1gp120 gene expression disruption in mycobacterium bovis BCG host strain: a critical issue for engineering mycobacterial based-vaccine vectors.
abstract: Mycobacterium bovis Bacillus Calmette-Guérin (BCG) as a live vector of recombinant bacterial vaccine is a promising system to be used. In this study, we evaluate the disrupted expression of heterologous HIV-1gp120 gene in BCG Pasteur host strain using replicative vectors pMV261 and pJH222. pJH222 carries a lysine complementing gene in BCG lysine auxotrophs. The HIV-1 gp120 gene expression was regulated by BCG hsp60 promoter (in plasmid pMV261) and Mycobacteria spp. alpha-antigen promoter (in plasmid pJH222). Among 14 rBCG:HIV-1gp120 (pMV261) colonies screened, 12 showed a partial deletion and two showed a complete deletion. However, deletion was not observed in all 10 rBCG:HIV-1gp120 (pJH222) colonies screened. In this study, we demonstrated that E. coli/Mycobacterial expression vectors bearing a weak promoter and lysine complementing gene in a recombinant lysine auxotroph of BCG could prevent genetic rearrangements and disruption of HIV 1gp120 gene expression, a key issue for engineering Mycobacterial based vaccine vectors.
authors.affiliation: AIDS Research Unit, Hospital Clínic/IDIBAPS-HIVACAT, University of Barcelona, 08036, Barcelona, Spain. jjoseph@clinic.ub.es
authors.forename: Joan
authors.initials: J
authors.lastname: Joseph
authors.forename: Raquel
authors.initials: R
authors.lastname: Fernández-Lloris
authors.forename: Elías
authors.initials: E
authors.lastname: Pezzat
authors.forename: Narcís
authors.initials: N
authors.lastname: Saubi
authors.forename: Pere-Joan
authors.initials: PJ
authors.lastname: Cardona
authors.forename: Beatriz
authors.initials: B
authors.lastname: Mothe
authors.forename: Josep Maria
authors.initials: JM
authors.lastname: Gatell
mesh_terms: D016915:AIDS Vaccines; D000595:Amino Acid Sequence; D001500:BCG Vaccine; D001483:Base Sequence; D015153:Blotting, Western; D017353:Gene Deletion; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009163:Mycobacterium bovis / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D014169:Transformation, Bacterial; D014614:Vaccines, Synthetic
chemical_list: D016915:AIDS Vaccines; D001500:BCG Vaccine; D015699:HIV Envelope Protein gp120; D014614:Vaccines, Synthetic; C514793:gp120 protein, Human immunodeficiency virus 1