A Baculovirus immediate-early gene, ie1, promoter drives efficient expression of a transgene in both Drosophila melanogaster and Bombyx mori

PLoS One. 2012;7(11):e49323. doi: 10.1371/journal.pone.0049323. Epub 2012 Nov 13.

Abstract

Many promoters have been used to drive expression of heterologous transgenes in insects. One major obstacle in the study of non-model insects is the dearth of useful promoters for analysis of gene function. Here, we investigated whether the promoter of the immediate-early gene, ie1, from the Bombyx mori nucleopolyhedrovirus (BmNPV) could be used to drive efficient transgene expression in a wide variety of insects. We used a piggyBac-based vector with a 3xP3-DsRed transformation marker to generate a reporter construct; this construct was used to determine the expression patterns driven by the BmNPV ie1 promoter; we performed a detailed investigation of the promoter in transgene expression pattern in Drosophila melanogaster and in B. mori. Drosophila and Bombyx belong to different insect orders (Diptera and Lepidoptera, respectively); however, and to our surprise, ie1 promoter-driven expression was evident in several tissues (e.g., prothoracic gland, midgut, and tracheole) in both insects. Furthermore, in both species, the ie1 promoter drove expression of the reporter gene from a relatively early embryonic stage, and strong ubiquitous ie1 promoter-driven expression continued throughout the larval, pupal, and adult stages by surface observation. Therefore, we suggest that the ie1 promoter can be used as an efficient expression driver in a diverse range of insect species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Baculoviridae / genetics*
  • Bombyx / embryology
  • Bombyx / genetics*
  • DNA / genetics
  • Drosophila melanogaster / genetics*
  • Embryo, Nonmammalian / metabolism
  • Gene Expression*
  • Genes, Immediate-Early / genetics*
  • Genes, Reporter
  • Genome, Insect / genetics
  • Green Fluorescent Proteins / metabolism
  • Larva / genetics
  • Mutagenesis, Insertional / genetics
  • Promoter Regions, Genetic*
  • Transformation, Genetic
  • Transgenes / genetics*

Substances

  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • DNA