Identification of a midgut-specific promoter in the silkworm Bombyx mori

Biochem Biophys Res Commun. 2013 Apr 19;433(4):542-6. doi: 10.1016/j.bbrc.2013.03.019. Epub 2013 Mar 21.

Abstract

The midgut is an important organ for digestion and absorption of nutrients and immune defense in the silkworm Bombyx mori. In an attempt to create a tool for midgut research, we cloned the 1080 bp P2 promoter sequence (P2P) of a highly expressed midgut-specific gene in the silkworm. The transgenic line (P2) was generated via embryo microinjection, in which the expression of EGFP was driven by P2P. There was strong green fluorescence only in the midgut of P2. RT-PCR and Western blot showed that P2P was a midgut-specific promoter with activity throughout the larval stage. A transgenic truncation experiment suggested that regions -305 to -214 and +107 to +181 were very important for P2P activity. The results of this study revealed that we have identified a midgut-specific promoter with a high level of activity in the silkworm that will aid future research and application of silkworm genes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Western
  • Bombyx / genetics*
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • DNA, Complementary / metabolism
  • Embryo, Nonmammalian / cytology
  • Embryo, Nonmammalian / metabolism
  • Fluorescence
  • Gastrointestinal Tract / cytology*
  • Genes, Insect*
  • Genetic Engineering / methods
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Larva / genetics
  • Larva / metabolism
  • Microinjections
  • Molecular Sequence Data
  • Organisms, Genetically Modified / genetics
  • Organisms, Genetically Modified / metabolism
  • Promoter Regions, Genetic*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Species Specificity

Substances

  • DNA, Complementary
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins