Use of a pooled clone method to isolate a novel Bacillus thuringiensis Cry2A toxin with activity against Ostrinia furnacalis

Shu, Changlong, Zhang, Jingtao, Chen, Guihua, Liang, Gemei, He, Kanglai, Crickmore, Neil, Huang, Dafang, Zhang, Jie and Song, Fuping (2013) Use of a pooled clone method to isolate a novel Bacillus thuringiensis Cry2A toxin with activity against Ostrinia furnacalis. Journal of Invertebrate Pathology, 114 (1). pp. 31-33. ISSN 0022-2011

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Abstract

A pooled clone method was developed to screen for cry2A genes. This metagenomic method avoids the need to analyse isolated Bacillus thuringiensis strains by performing gene specific PCR on plasmid-enriched DNA prepared from a pooled soil sample. Using this approach the novel holotype gene cry2Ah1 was cloned and characterized. The toxin gene was over-expressed in Escherichia coli Rosetta (DE3) and the expressed toxin accumulated in both the soluble and insoluble fractions. The soluble Cry2Ah1 was found to have a weight loss activity against Ostrinia furnacalis, and a growth inhibitory activity to both Cry1Ac-susceptible and resistant Helicoverpa armigera populations.

Item Type: Article
Schools and Departments: School of Life Sciences > Biochemistry
Subjects: Q Science > QR Microbiology > QR0075 Bacteria
Depositing User: Neil Crickmore
Date Deposited: 05 Sep 2014 14:07
Last Modified: 05 Sep 2014 14:07
URI: http://sro.sussex.ac.uk/id/eprint/49728
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