A novel recombinant baculovirus overexpressing a Bacillus thuringiensis Cry1Ab toxin enhances insecticidal activityReport as inadecuate




A novel recombinant baculovirus overexpressing a Bacillus thuringiensis Cry1Ab toxin enhances insecticidal activity - Download this document for free, or read online. Document in PDF available to download.

Biological Procedures Online

, 16:7

First Online: 15 April 2014Received: 29 January 2014Accepted: 29 March 2014DOI: 10.1186-1480-9222-16-7

Cite this article as: El-Menofy, W., Osman, G., Assaeedi, A. et al. Biol Proced Online 2014 16: 7. doi:10.1186-1480-9222-16-7

Abstract

Baculoviruses have been genetically modified to express foreign genes under powerful promoters in order to accelerate their speed of killing. In this study a truncated form of cry1Ab gene derived from Bacillus thuringinsis Bt subsp. aegypti isolate Bt7 was engineered into the genome of the baculovirus Autographa californica multiple nuclearpolyhedrosis wild type virus, in place of the polyhedrin gene by using homologous recombination in Spodoptera frugiperda Sf cells between a transfer vector carrying the Bt gene and the wild type virus linearized DNA. Recombinant wild type virus containing the cry1Ab gene was detected as blue occlusion-negative plaques in monolayers of Sf cells grown in the presence of X-Gal. In Sf cells infected with plaque-purified recombinant virus, the cry1Ab gene was expressed to yield a protein of approximately 82-kDa, as determined by immunoblot analysis. The toxicity of the recombinant virus expressing the insecticidal crystal protein ICP was compared to that of the wild-type virus. Infected-cell extract was toxic to cotton leaf worm Spodoptera littoralis second instar larvae and the estimated LC50 was 1.7 μg-ml for the recombinant virus compared with that of wild-type virus which was 10 μg-ml.

KeywordsBacillus thuringiensis Toxin Baculovirus Cry 1Ab LC50 Electronic supplementary materialThe online version of this article doi:10.1186-1480-9222-16-7 contains supplementary material, which is available to authorized users.

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Author: Wael El-Menofy - Gamal Osman - Abdulrahman Assaeedi - Mohamed Salama

Source: https://link.springer.com/







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