Expression of mosquito-larvicidal toxin genes under the control of a native promoter in Enterobacter amnigenus An11

Biotechnol Lett. 2013 Aug;35(8):1331-7. doi: 10.1007/s10529-013-1209-3. Epub 2013 Apr 23.

Abstract

Enterobacter amnigenus An11, that can colonize the gut of mosquito larva, is an alternative toxin-producing host to be used as a mosquito control since it is able to float in the feeding zone of mosquito larvae. To produce mosquito-larvicidal toxins in this bacterium, a native promoter has been identified from its genomic DNA. The promoter exhibited consensus sequences for -35 and -10 regions of bacterial promoters and constitutively drove the expression of gfp. This promoter was inserted into recombinant plasmids upstream of promoter-free cyt2Aa2 from Bacillus thuringiensis and mtx2 from Bacillus sphaericus. Results demonstrated that Cyt2Aa2 and Mtx2 are constitutively produced without induction. The recombinant E. amnigenus showed toxicity against mosquito larvae, demonstrating a potential to be applied in a mosquito control program.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / biosynthesis*
  • Bacterial Proteins / genetics*
  • Culicidae / growth & development
  • Culicidae / microbiology
  • Culicidae / physiology
  • Endotoxins / biosynthesis*
  • Endotoxins / genetics*
  • Enterobacter / enzymology*
  • Enterobacter / genetics*
  • Gene Expression*
  • Genetic Vectors
  • Hemolysin Proteins / biosynthesis*
  • Hemolysin Proteins / genetics*
  • Larva / microbiology
  • Larva / physiology
  • Metabolic Engineering / methods
  • Mosquito Control / methods
  • Pest Control, Biological / methods
  • Plasmids
  • Promoter Regions, Genetic
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Survival Analysis

Substances

  • Bacillus thuringiensis Toxins
  • Bacterial Proteins
  • Endotoxins
  • Hemolysin Proteins
  • Recombinant Proteins
  • insecticidal crystal protein, Bacillus Thuringiensis