[Construction of insecticidal recombinant Bacillus thuringiensis using an integrative vector]

Yi Chuan Xue Bao. 2003 Aug;30(8):737-42.
[Article in Chinese]

Abstract

Insecticidal crystal protein gene cry1C which is highly toxic to Spodoptera exigua was cloned into an integrative vector pBMB-F7E, which was derived from Bacillus thuringiensis transposon Tn4430. The recombinant integrative plasmid pBMB-FLC harboring the cry1C gene was gained and transformed into a wild-type Bt strain YBT803-1. A transformant BMB803-A was obtained, and grown at 46 degrees C for about 120 generations, From which three recombinants with cry1C gene integrating into the chromosome were achieved at a frequency of approximately 3.4 x 10(-5). Southern blotting revealed that the integration occurred in different sites of the chromosome. The integrated cry1C were expressed effectively. The results of bioassays showed that the toxicity of recombinants BMB803-X and BMB803-Z against Plutella xylostella were similar to that of strain YBT803-1, and their toxicity to Spodoptera exigua were higher than that of strain YBT803-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacillus thuringiensis / genetics*
  • Bacillus thuringiensis / growth & development
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Bacterial Toxins*
  • DNA, Bacterial / genetics
  • DNA, Bacterial / metabolism
  • Deoxyribonucleases, Type II Site-Specific / metabolism
  • Endotoxins / genetics*
  • Endotoxins / metabolism
  • Genetic Vectors / genetics*
  • Hemolysin Proteins
  • Microscopy, Electron
  • Pest Control, Biological / methods
  • Spodoptera / microbiology
  • Spores, Bacterial / genetics
  • Spores, Bacterial / ultrastructure
  • Transformation, Genetic

Substances

  • Bacillus thuringiensis Toxins
  • Bacterial Proteins
  • Bacterial Toxins
  • DNA, Bacterial
  • Endotoxins
  • Hemolysin Proteins
  • insecticidal crystal protein, Bacillus Thuringiensis
  • ATGCAT-specific type II deoxyribonucleases
  • Deoxyribonucleases, Type II Site-Specific