Comparative Expression of the Hepatitis B Surface Antigen Gene in Biochemically Transformed Human, Simian and Murine Cells

Colbère-Garapin, Florence; Horaud, Florian; Kourilsky, Philippe; Garapin, Axel-Claude

doi:10.1099/0022-1317-66-8-1741

Cited by 7 publications

(1 citation statement)

References 34 publications

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“…Transfection. Plasmids containing PV-1 full-length infectious cDNA were used to transfect Vero cell monolayers at about 80% confluence in 30-mm-diameter dishes by the calcium phosphate technique (8,22). Briefly, medium was removed from dishes, and cells were transfected with about 5 p,g of plasmid DNA in 0.5 ml of calcium phosphate precipitate in the presence of 10 ,ug of salmon sperm carrier DNA and 0.17 mg of aprotinin (Sigma A6279) in N- 2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)buffered saline (52).…”

Section: Methodsmentioning

confidence: 99%

Molecular characterization of mouse-virulent poliovirus type 1 Mahoney mutants: involvement of residues of polypeptides VP1 and VP2 located on the inner surface of the capsid protein shell

Couderc

Hogle

Blay

et al. 1993

J Virol

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Using an in vivo approach, we isolated two mouse-neurovirulent PV-1 mutants in the mouse central nervous system after a single passage of PV-1/Mahoney inoculated by the intracerebral route. Both mutants were subjected to two additional passages in mice, plaque purified, and subsequently characterized. The two cloned mutants, Mah-NK13 and Mah-NL32, retained phenotypic characteristics of the parental PV-1/Mahoney, including epitope map, heat lability, and temperature sensitivity. Mah-NK13 exhibited slightly smaller plaques than did the parental virus. The nucleotide sequences of the mutant genomes were determined, and mutations were identified. Mutations were independently introduced into the parental PV-1/Mahoney genome by single-site mutagenesis. Mutated PV-1/Mahoney viruses were then tested for their neurovirulence in mice. A single amino acid substitution in the capsid proteins VP1 (Thr-22-*Ile) and VP2 (Ser-31-Thr) identified in the Mah-NK13 and Mah-NL32 genomes, respectively, conferred the mouse-virulent phenotype to the mouse-avirulent PV-1/Mahoney. Ile-22 in VP1 was responsible for the small-plaque phenotype of Mah-NK13. Both mutations arose during the first passage in the mouse central nervous system. We thus identified a new mouse adaptation determinant on capsid protein VP1, and we showed that at least one other capsid protein, VP2, could also express a mouse adaptation determinant. Both determinants are located in the inside of the three-dimensional structure of the viral capsid. They may be involved in the early steps of mouse nerve cell infection subsequent to receptor attachment.

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Section: Methodsmentioning

confidence: 99%