Expression of cry1Ab gene from a novel Bacillus thuringiensis strain SY49-1 active on pest insects

Braz J Microbiol. 2016 Jul-Sep;47(3):597-602. doi: 10.1016/j.bjm.2016.04.011. Epub 2016 Apr 20.

Abstract

In this study, the cry1Ab gene of previously characterized and Lepidoptera-, Diptera-, and Coleoptera-active Bacillus thuringiensis SY49-1 strain was cloned, expressed and individually tested on Ephestia kuehniella (Lepidoptera: Pyralidae) and Plodia interpunctella (Lepidoptera: Pyralidae) larvae. pET-cry1Ab plasmids were constructed by ligating the cry1Ab into pET28a (+) expression vector. Constructed plasmids were transferred to an Escherichia coli BL21 (DE3) strain rendered competent with CaCl2. Isopropyl β-d-1-thiogalactopyranoside was used to induce the expression of cry1Ab in E. coli BL21(DE3), and consequently, ∼130kDa of Cry1Ab was obtained. Bioassay results indicated that recombinant Cry1Ab at a dose of 1000μgg(-1) caused 40% and 64% mortality on P. interpunctella and E. kuehniella larvae, respectively. However, the mortality rates of Bt SY49-1 strains' spore-crystal mixture at the same dose were observed to be 70% on P. interpunctella and 90% on E. kuehniella larvae. The results indicated that cry1Ab may be considered as a good candidate in transgenic crop production and as an alternative biocontrol agent in controlling stored product moths.

Keywords: Bt SY49-1; Gene cloning; Insecticidal activity; Recombinant Cry1Ab.

MeSH terms

  • Animals
  • Bacillus thuringiensis / physiology*
  • Bacillus thuringiensis / ultrastructure
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Bacterial Proteins / toxicity
  • Cloning, Molecular
  • Endotoxins / genetics*
  • Endotoxins / metabolism
  • Endotoxins / toxicity
  • Gene Expression*
  • Hemolysin Proteins / genetics*
  • Hemolysin Proteins / metabolism
  • Hemolysin Proteins / toxicity
  • Insect Control* / methods
  • Insecticides
  • Larva
  • Moths / drug effects

Substances

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