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Applied and Environmental Microbiology, February 2005, p. 761-765, Vol. 71, No. 2
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.2.761-765.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

New Strategy for Identification of Novel cry-Type Genes from Bacillus thuringiensis Strains

Corina M. Berón, Leonardo Curatti,{dagger} and Graciela L. Salerno*

Centro de Investigaciones Biológicas, Fundación para Investigaciones Biológicas Aplicadas, Mar del Plata, Argentina

Received 22 May 2004/ Accepted 15 September 2004

We designed five degenerate primers for detection of novel cry genes from Bacillus thuringiensis strains. An efficient strategy was developed based on a two-step PCR approach with these primers in five pair combinations. In the first step, only one of the primer pairs is used in the PCR, which allows amplification of DNA fragments encoding protein regions that include consensus domains of representative proteins belonging to different Cry groups. A second PCR is performed by using the first-step amplification products as DNA templates and the set of five primer combinations. Cloning and sequencing of the last-step amplicons allow both the identification of known cry genes encoding Cry proteins covering a wide phylogenetic distance and the detection and characterization of cry-related sequences from novel B. thuringiensis isolates.

* Corresponding author. Mailing address: Centro de Investigaciones Biológicas, Fundación para Investigaciones Biológicas Aplicadas (FIBA), C.C. 1348, 7600 Mar del Plata, Argentina. Phone: 54-223-4748257. Fax: 54-223-4757120. E-mail: gsalerno{at}fiba.org.ar.

{dagger} Present address: Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720-3102.

Applied and Environmental Microbiology, February 2005, p. 761-765, Vol. 71, No. 2
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.2.761-765.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Guo, S., Liu, M., Peng, D., Ji, S., Wang, P., Yu, Z., Sun, M. (2008). New Strategy for Isolating Novel Nematicidal Crystal Protein Genes from Bacillus thuringiensis Strain YBT-1518. Appl. Environ. Microbiol. 74: 6997-7001 [Abstract] [Full Text]  


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