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Identification and characterization of novel vip3-like genes in Bacillus thuringiensis Straits from a Spanish collection

AuthorsPalma, Leopoldo ; Maeztu, Mireya; Ruiz de Escudero, Íñigo ; Muñoz, Delia ; Caballero, Primitivo
Issue DateJun-2011
PublisherInternational Organisation for Biological and Integrated Control of Noxious Animals and Plants
CitationIOBC/WPRS Bulletin 66: 75-78 (2011)
AbstractA total of 400 strains of Bacillus thuringiensis (Bt) have been screened to identify the presence of vip3-like genes using a PCR-restriction fragment length polymorphism (PCR-RFLP) strategy to detect novel vip3 genes. This allowed identification of vip3A known genes in ~80% of the strains, whereas three new RFLP patterns revealed the presence of three novel vip3-like genes in three different Bt strains (B5.3, B8.1, B8.3). PCR amplicons from B8.1 and B8.3 strains produced fragments of 1646 and 1642 bp, respectively that were cloned and sequenced. These two fragments shared homologies of ~77% with known vip3 genes suggesting they may be novel vip3 genes. The strain B5.3 amplicon was 1620 bp and showed 91% homology with vip3Aa1 gene. The full-length sequence of this vip3A-like gene was obtained and found to be a new vip3Ag variant and was designated vip3Ag4 by the Bt Toxin Nomenclature Committee. This gene is 2364 bp in size, encodes a 788 amino acid (88 kDa) protein and shares between 81 and 91% homology with known vip3A genes. Furthermore, vip3Ag4 was successfully cloned in pET-28b(+) and expressed in Escherichia coli BL21(DE3). The protein obtained was toxic against Chrysodeixis chalcites, Helicoverpa armigera, Spodoptera exigua, S. frugiperda and S. littoralis.
DescriptionTrabajo presentado en el 13th European Meeting Biological Control in IPM Systems, celebrado en Innsbruck (Austria) del 19 al 23 de junio de 2011.
Appears in Collections:(IDAB) Libros y partes de libros
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