Development and characterization of a double subgenomic chikungunya virus infectious clone to express heterologous genes in Aedes aegypti mosquitoes
Autor(es): Vanlandingham Dana L, Tsetsarkin Konstantin, Hong Chao, Klingler Kimberly, McElroy Kate L, Lehane Michael J, Higgs Stephen
Resumo: Three full-length infectious cDNA clones based on the alphavirus chikungunya (CHIKV) were developed - characterized in vitro - in vivo. The full-length clone retained the viral phenotypes of CHIKV in both cell culture - in mosquitoes - should be a valuable tool for the study of virus interactions in an epidemiologically significant natural vector, Aedes aegypti. Two additional infectious clones were constructed that express green fluorescent protein (EGFP) in the midgut, salivary gl-s, - nervous tissue of Aedes aegypti mosquitoes following oral infection. The two constructs differed in the placement of the subgenomic promoter - the gene encoding EGFP. Viruses derived from the pCHIKic EGFP constructs (5' CHIKV EGFP - 3' CHIKV EGFP) expressed EGFP in 100% of the Ae. aegypti mosquitoes tested on days 7 - 14 post infection (p.i.). The 5' CHIKV EGFP disseminated to 90% of the salivary gl-s - nervous tissue by day 14 p.i. Dissemination rates of this new viral vector exceeds those of previous systems, thus exp-ing the repertoire - potential for gene expression studies on this important vector species.
Palavras-Chave: Mosquitoes; Aedes aegypti; Chikungunya virus; Green fluorescent protein; Gene expression
Imprenta: Insect Biochemistry and Molecular Biology, v. 35, n. 10, p. 1162-1170, 2005
Identificador do objeto digital: 10.1016/j.ibmb.2005.05.008
Descritores: Chikungunya virus - Cell ; Chikungunya virus - Cytopathology ; Chikungunya virus - Genome ; Chikungunya virus - Pathogenesis ; Chikungunya virus - Proteins ; Chikungunya Virus - Virus
Data de publicação: 2005
