Dengue fever is one of the most rapidly expanding diseases in the tropics with over 2 billion people at risk. Transmission of Dengue virus (DEN) involves a complex interaction between the genomes of the primary mosquito vector, Aedes aegypti, the virus, and its human host. The proposed research will focus on two of these components by examiningthe genetics of vector competence in Ae. aegypti and the degree to which vector competence is affected by genetic variance in the DEN virus. We have mapped Quantitative Trait Loci (QTL) affecting the ability of Ae. aegypri midgut to become infected with Dengue-2 virus (DEN2). These twomidgut infection barrier (MIB) QTL are called dmib2, dmib3 and were mapped to chromosomes II and III respectively. The primary goal of the proposed work is to use marker-assisted selection (MAS) to generate 4 strains of Ae. aegypri. These will be a strain homozygous susceptible at both loci (DS2), a strain homozygous refractory at both loci (DR2) and 2 strains homozygous susceptible at one locus and homozygous refractory at the other locus (DMIB2, DMIB3). These strains will provide an exceptional opportunity to elucidate the phenotypes associated with each genotype at the QTL underlying DEN transmission. DS2 will be used to isolate and map DEN barrier QTL from worldwide populations of Ae. aegypri to determine if additional QTL control DEN susceptibility and/or if alternate alleles exist at the currently identified QTL. DMIB2, DMIB3, and DR2 will be used to assess how alleles at both loci separately and together confer susceptibility in different DEN serotypes and genotypes in each of the 4 serotypes. DS2 and DR2 will be used in the construction of recombinant inbred lines for use in a subtractive hybridization approach to clone cDNAs putatively encoded bydmib3and to assist in a collaborative effort to positionally clone dmib3.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI049256-01
Application #
6317871
Study Section
Special Emphasis Panel (ZRG1-TMP (01))
Program Officer
Aultman, Kathryn S
Project Start
2001-04-01
Project End
2006-03-31
Budget Start
2001-04-01
Budget End
2002-03-31
Support Year
1
Fiscal Year
2001
Total Cost
$313,999
Indirect Cost
Name
Colorado State University-Fort Collins
Department
Microbiology/Immun/Virology
Type
Schools of Veterinary Medicine
DUNS #
112617480
City
Fort Collins
State
CO
Country
United States
Zip Code
80523
Lozano-Fuentes, Saul; Fernandez-Salas, Ildefonso; de Lourdes Munoz, Maria et al. (2009) The neovolcanic axis is a barrier to gene flow among Aedes aegypti populations in Mexico that differ in vector competence for Dengue 2 virus. PLoS Negl Trop Dis 3:e468
Black 4th, William C; Gorrochetegui-Escalante, Norma; Randle, Nadine P et al. (2008) The Yin and Yang of linkage disequilibrium: mapping of genes and nucleotides conferring insecticide resistance in insect disease vectors. Adv Exp Med Biol 627:71-83
Black 4th, W C; Vontas, J G (2007) Affordable assays for genotyping single nucleotide polymorphisms in insects. Insect Mol Biol 16:377-87
Black 4th, W C; Gorrochotegui-Escalante, N; Duteau, N M (2006) Heated oligonucleotide ligation assay (HOLA): an affordable single nucleotide polymorphism assay. J Med Entomol 43:238-47
Richardson, Jason; Molina-Cruz, Alvaro; Salazar, Ma Isabel et al. (2006) Quantitative analysis of dengue-2 virus RNA during the extrinsic incubation period in individual Aedes aegypti. Am J Trop Med Hyg 74:132-41
Gorrochotegui-Escalante, Norma; Lozano-Fuentes, Saul; Bennett, Kristine E et al. (2005) Association mapping of segregating sites in the early trypsin gene and susceptibility to dengue-2 virus in the mosquito Aedes aegypti. Insect Biochem Mol Biol 35:771-88
Bennett, Kristine E; Beaty, Barry J; Black 4th, William C (2005) Selection of D2S3, an Aedes aegypti (Diptera: Culicidae) strain with high oral susceptibility to Dengue 2 virus and D2MEB, a strain with a midgut barrier to Dengue 2 escape. J Med Entomol 42:110-9
Bennett, Kristine E; Flick, Don; Fleming, Karen H et al. (2005) Quantitative trait loci that control dengue-2 virus dissemination in the mosquito Aedes aegypti. Genetics 170:185-94
Gomez-Machorro, C; Bennett, K E; del Lourdes Munoz, M et al. (2004) Quantitative trait loci affecting dengue midgut infection barriers in an advanced intercross line of Aedes aegypti. Insect Mol Biol 13:637-48
Gorrochotegui-Escalante, N; Black 4th, W C (2003) Amplifying whole insect genomes with multiple displacement amplification. Insect Mol Biol 12:195-200

Showing the most recent 10 out of 11 publications