Related Experiment Videos

Expression of 135-kDa insecticidal protein gene from Bacillus thuringiensis in the yeast Saccharomyces cerevisiae

K Watanabe1, M Toyoda, H Hashimoto

  • 1Showa Woman's University, Tokyo, Japan.

Bioscience, Biotechnology, and Biochemistry
|September 1, 1996
PubMed

Insights

Bacillus thuringiensis insecticidal protein production in yeast was achieved by modifying the cryIA(a) gene. This engineered yeast demonstrated efficacy against Plutella xylostella larvae.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Entomology

Background:

  • Bacillus thuringiensis subsp. aizawai synthesizes CryIA(a) insecticidal proteins.
  • Gene expression in Saccharomyces cerevisiae can be challenging due to regulatory elements.

Purpose of the Study:

  • To enable heterologous expression of Bacillus thuringiensis cryIA(a) gene in yeast.
  • To investigate the insecticidal properties of yeast-expressed CryIA(a) protein.

Main Methods:

  • Transformation of Saccharomyces cerevisiae with a cryIA(a) gene vector.
  • Modification of the cryIA(a) gene by removing the 5'-upstream region.
  • Assaying insecticidal activity against Plutella xylostella.

Main Results:

  • Initial transformation did not yield observable CryIA(a) protein expression.
  • Removing the 5'-upstream region enabled successful CryIA(a) protein synthesis in yeast.
  • Yeast extract containing CryIA(a) protein exhibited insecticidal activity against Plutella xylostella.

Conclusions:

  • The 5'-upstream region of the cryIA(a) gene inhibits its expression in Saccharomyces cerevisiae.
  • Genetic modification of Bacillus thuringiensis genes can facilitate their expression in yeast for pest control applications.

Related Concept Videos