Development of a Sindbis virus expression system that efficiently expresses green fluorescent protein in midguts of Aedes aegypti following per os infection.
A double subgenomic Sindbis (dsSIN) virus, MRE/3'2 J/GFP, was constructed to efficiently express green fluorescent protein (GFP) in the midgut of Aedes aegypti following per os infection. The MRE/3'2 J/GFP RNA genome contained the nonstructural genes and cis-acting sequences of the dsSIN virus, TE/3'2 J/GFP, but had the structural genes of MRE16 SIN virus. MRE/3'2 J/GFP virus, unlike TE/3'2 J/GFP virus, efficiently infected mosquitoes orally. At 1-2 days postinfection, GFP was observed as multiple foci of expression on the lumenal side of the midgut. At 10-12 days postinfection, thirteen of fifteen mosquitoes infected with MRE/3'2 J/GFP virus had high levels of GFP expression in the mosquito midgut. The MRE3'2 J dsSIN expression system should be an important tool for efficient gene expression in Ae. aegypti midguts.
Olson
K E
KE
Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Colorado State University, Ft. Collins, CO 80523, USA. kolson@cvmbs.colostate.edu
Myles
K M
KM
Seabaugh
R C
RC
Higgs
S
S
Carlson
J O
JO
Beaty
B J
BJ
D000818:Animals; D002478:Cells, Cultured; D003465:Culex / Q000235:genetics / Q000821:virology*; D004064:Digestive System; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics*; D012845:Sindbis Virus / Q000235:genetics*; D017361:Viral Nonstructural Proteins / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics; D014773:Virology / Q000379:methods*
D008164:Luminescent Proteins; D017361:Viral Nonstructural Proteins; D015678:Viral Structural Proteins; D049452:Green Fluorescent Proteins