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J Gen Virol 88 (2007), 1545-1554; DOI 10.1099/vir.0.82577-0

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Genetic determinants of Sindbis virus strain TR339 affecting midgut infection in the mosquito Aedes aegypti

Dennis J. Pierro, Erik L. Powers and Ken E. Olson

Arthropod-Borne and Infectious Disease Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523, USA

Correspondence
Ken E. Olson
kolson{at}colostate.edu

Mosquito midgut epithelial cells (MEC) play a major role in determining whether an arbovirus can successfully infect and be transmitted by mosquitoes. The Sindbis virus (SINV) strain TR339 efficiently infects Aedes aegypti MEC but the SINV strain TE/5'2J poorly infects MEC. SINV determinants for MEC infection have been localized to the E2 glycoprotein. The E2 amino acid sequences of TR339 and TE/5'2J differ at two sites, E2-55 and E2-70. We have altered the TE/5'2J virus genome by site-directed mutagenesis to contain two TR339 residues, E2-55 H->Q (histidine to glutamine) and E2-70 K->E (lysine to glutamic acid). We have characterized the growth patterns of derived viruses in cell culture and determined the midgut infection rate (MIR) in A. aegypti mosquitoes. Our results clearly show that the E2-55 H->Q and the E2-70 K->E mutations in the TE/5'2J virus increase MIR both independently and in combination. TE/5'2J virus containing both TR339 E2 residues had MIRs similar to the parental TR339 virus. In addition, SINV propagated in a mammalian cell line had a significantly lower A. aegypti midgut 50 % infectious dose than virus propagated in a mosquito cell line.




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D. J. Pierro, E. L. Powers, and K. E. Olson
Genetic Determinants of Sindbis Virus Mosquito Infection Are Associated with a Highly Conserved Alphavirus and Flavivirus Envelope Sequence
J. Virol., March 15, 2008; 82(6): 2966 - 2974.
[Abstract] [Full Text] [PDF]




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