Infectious clone construction of dengue virus type 2, strain Jamaican 1409, and characterization of a conditional E6 mutation
Autor(es): Pierro, Dennis J.; Salazar, Ma Isabel; Beaty, Barry J.; Olson, Ken E.
Resumo: A full-length infectious cDNA clone (ic) was constructed from the genome of the dengue virus type 2 (DENV-2) Jamaica83 1409 strain, pBAC1 409ic,by using a bacterial artifical chromosome plasmid system. Infectious virus was generated and characterized for growth in cell culture and for infection in Aedes aegypti mosquitoes. During construction, an isoleucine to methionine (lle -> Met) change was found at position 6 in the envelope glycoprotein Sequence between low- and high-passage DENV-2 1409 strains. In vitro-transcribed genomic RNA of 1409ic with E6-lle produced infectious virions following electroporation in mosquito cells, but not mammalian cells, while 1409ic RNA with an E6-Met mutation produced virus in both cell types. Moreover, DENV-2 1409 with the E6-lle residue produced syncytia in C6/36 cell culture, whereas viruses with E6-Met did not. However, in vitro cell culture-derived growth-curve data and in vivo mosquito-infection rates revealed that none of the analysed DENV-2 strains differed from each other.
Palavras-Chave: Borne Encephalitis-Virus; Amino-Acid-Sequence; Envelope Glycoprotein; Aedes-Aegypti; Phylogenetic-Relationships; Nucleotide-Sequence; Vector Competence; Membrane-Fusion; Cell-Cultures; Cdna-Clones
Imprenta: Journal of General Virology, v. 87, p. 2263-2268, 2006
Identificador do objeto digital: 10.1099/vir.0.81958-0
Descritores: Aedes aegypti - Biochemistry ; Aedes aegypti - Cell ; Aedes aegypti - DNA ; Aedes aegypti - Genome ; Aedes aegypti - RNA ; Aedes aegypti - Infectious diseases ; Aedes aegypti - Virus
Data de publicação: 2006
