Expression of the chloramphenicol acetyltransferase gene in Aedes albopictus (C6/36) cells using a non-infectious Sindbis virus expression vector

Insect Mol Biol. 1992;1(1):49-52. doi: 10.1111/j.1365-2583.1993.tb00077.x.

Abstract

Genomic RNA was transcribed in vitro from the non-infectious Sindbis (SIN) virus expression vector (pTRCAT) and introduced into C6/36 (Aedes albopictus) cells by liposome-mediated transfections. The chloramphenicol acetyltransferase (CAT) polypeptide expressed within cells was detected by an indirect immunofluorescent assay directly into 24-well polystyrene tissue culture plates. Approximately 1 in 1000 C6/36 cells showed fluorescence when the transfection was optimized. CAT enzyme activity was also assayed; in C6/36 cells CAT expression was detected as early as 8 h after transfection, peaked at 24 h, and could still be detected at 7 days. At 24 h posttransfection each transfected C6/36 cell was calculated to express 1.3 x 10(7) CAT polypeptides.

Publication types

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

MeSH terms

  • Aedes / enzymology
  • Aedes / genetics*
  • Animals
  • Cell Line
  • Chick Embryo
  • Chloramphenicol O-Acetyltransferase / biosynthesis
  • Chloramphenicol O-Acetyltransferase / genetics*
  • Gene Expression Regulation
  • Genetic Vectors
  • Sindbis Virus / genetics*
  • Transfection

Substances

  • Chloramphenicol O-Acetyltransferase