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Journal of Virology, March 2008, p. 2966-2974, Vol. 82, No. 6
0022-538X/08/$08.00+0     doi:10.1128/JVI.02060-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Genetic Determinants of Sindbis Virus Mosquito Infection Are Associated with a Highly Conserved Alphavirus and Flavivirus Envelope Sequence{triangledown}

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, Colorado 80523

Received 17 September 2007/ Accepted 15 December 2007

Wild-type Sindbis virus (SINV) strain MRE16 efficiently infects Aedes aegypti midgut epithelial cells (MEC), but laboratory-derived neurovirulent SINV strain TE/5'2J infects MEC poorly. SINV determinants for MEC infection have been localized to the E2 glycoprotein. The E2 amino acid sequences of MRE16 and TE/5'2J differ at 60 residue sites. To identify the genetic determinants of MEC infection of MRE16, the TE/5'2J virus genome was altered to contain either domain chimeras or more focused nucleotide substitutions of MRE16. The growth patterns of derived viruses in cell culture were determined, as were the midgut infection rates (MIR) in A. aegypti mosquitoes. The results showed that substitutions of MRE16 E2 aa 95 to 96 and 116 to 119 into the TE/5'2J virus increased MIR both independently and in combination with each other. In addition, a unique PPF/.GDS amino acid motif was located between these two sites that was found to be a highly conserved sequence among alphaviruses and flaviviruses but not other arboviruses.

* Corresponding author. Present address: Room 6132, Laboratory of Infectious Diseases, NIAID-NIH, 50 South Drive, Bethesda, MD 20892. Phone: (301) 594-2271. Fax: (301) 496-8312. E-mail: pierrod{at}niaid.nih.gov

{triangledown} Published ahead of print on 26 December 2007.

Journal of Virology, March 2008, p. 2966-2974, Vol. 82, No. 6
0022-538X/08/$08.00+0     doi:10.1128/JVI.02060-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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