Development of a Sindbis virus expression system that efficiently expresses green fluorescent protein in midguts of Aedes aegypti following per os infection

Insect Mol Biol. 2000 Feb;9(1):57-65. doi: 10.1046/j.1365-2583.2000.00162.x.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cells, Cultured
  • Culex / genetics
  • Culex / virology*
  • Digestive System
  • Gene Expression Regulation, Viral*
  • Genes, Viral
  • Green Fluorescent Proteins
  • Luminescent Proteins / genetics*
  • Sindbis Virus / genetics*
  • Viral Nonstructural Proteins / genetics
  • Viral Structural Proteins / genetics
  • Virology / methods*

Substances

  • Luminescent Proteins
  • Viral Nonstructural Proteins
  • Viral Structural Proteins
  • Green Fluorescent Proteins