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Multiplex PCR screening to detect cry9 genes in Bacillus thuringiensis strains
E Ben-Dov1, Q Wang, A Zaritsky
1Department of Life Sciences, Ben-Gurion University of the Negev, Be'er-Sheva 84105, Israel.
Applied and Environmental Microbiology
|July 31, 1999
Summary
Researchers developed new PCR primers to identify Bacillus thuringiensis (Bt) strains carrying specific crystal protein (cry) genes. This advancement aids in classifying Bt strains for pest control applications.
Area of Science:
- Microbiology
- Molecular Biology
- Entomology
Background:
- Bacillus thuringiensis (Bt) is a bacterium used for biological pest control.
- Bt produces crystal proteins (Cry) toxic to specific insect pests.
- Existing PCR methods need optimization for comprehensive Cry gene identification.
Purpose of the Study:
- To design and validate a new set of PCR primers for identifying cry9 genes in Bt.
- To complement existing PCR methodologies for broader cry gene detection.
- To rapidly screen and classify Bt strains based on cry gene profiles.
Main Methods:
- Design of universal (Un9) and specific multiplex PCR primers for cry9 genes.
- PCR analysis of 16 standard Bt strains and 215 field isolates.
- Classification of isolates based on cry9 gene presence and profile.
Main Results:
- The new primers successfully screened Bt strains for cry9 genes.
- B. thuringiensis subsp. aizawai HD-133 tested positive with Un9 but negative for cry9 genes.
- 22 field isolates were positive with Un9 and classified into three cry9 profiles, all containing cry1 and cry2 genes.
Conclusions:
- The developed primer set enhances the identification and classification of Bt strains.
- This methodology aids in understanding the diversity of cry genes in Bt.
- The primers are valuable tools for Bt research and application in pest management.