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Infectious clone construction of dengue virus type 2, strain Jamaican 1409, and characterization of a conditional E6 mutation

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

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

A full-length infectious cDNA clone (ic) was constructed from the genome of the dengue virus type 2 (DENV-2) Jamaica83 1409 strain, pBAC1409ic, 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 (IleMet) 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-Ile 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-Ile 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.

This article has been cited by other articles:

				D. E. Brackney, B. D. Foy, and K. E. Olson The Effects of Midgut Serine Proteases on Dengue Virus Type 2 Infectivity of Aedes aegypti Am J Trop Med Hyg, August 1, 2008; 79(2): 267 - 274. [Abstract] [Full Text] [PDF]