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PCR-based approach for detection of novel Bacillus thuringiensis cry genes

Juarez-Perez Victor-Manuel, Ferrandis M.D., Frutos Roger. 1997. PCR-based approach for detection of novel Bacillus thuringiensis cry genes. Applied and Environmental Microbiology, 63 (8) : 2997-3002.

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Autre titre : Détection de nouveaux gènes Cry de Bacillus thuringiensis par une approche à l'aide de la technique PCR

Résumé : A two-step strategy, named exclusive PCR or E-PCR, has been developed to overcome the main limitation of PCR, which is the detection of already-known sequences only. This strategy allows the ability to detect and further clone and sequence genes for which no specific primers are available and in which a variable region exists between two conserved regions. This approach has been applied to #Bacillus thuringiensis #cryI# genes by the use of mixtures of degenerate and specific primers recognizing well-known sequences. The first step allows the accurate identification of already-characterized #cryI# genes by the use of three primers. During the second step, the same sets of primers are used to exclude known sequences and to positively detect #cryI# genes unrecognized by any specific primer. The method, as well as its application to detect, clone, and sequence a novel #cryI# gene, is described in this article.

Mots-clés Agrovoc : Bacillus thuringiensis, identification, gène, PCR, technique analytique

Classification Agris : H20 - Maladies des plantes
U30 - Méthodes de recherche

Auteurs et affiliations

  • Juarez-Perez Victor-Manuel, CIRAD-GERDAT-BIOTROP/IGEPAM (FRA)
  • Ferrandis M.D.
  • Frutos Roger, CIRAD-GERDAT-BIOTROP/IGEPAM (FRA)

Autres liens de la publication

Source : Cirad - Agritrop (https://agritrop.cirad.fr/401070/)

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