title: Use of a pooled clone method to isolate a novel Bacillus thuringiensis Cry2A toxin with activity against Ostrinia furnacalis.
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.
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, PR China.
authors.forename: Changlong
authors.initials: C
authors.lastname: Shu
authors.forename: Jingtao
authors.initials: J
authors.lastname: Zhang
authors.forename: Guihua
authors.initials: G
authors.lastname: Chen
authors.forename: Gemei
authors.initials: G
authors.lastname: Liang
authors.forename: Kanglai
authors.initials: K
authors.lastname: He
authors.forename: Neil
authors.initials: N
authors.lastname: Crickmore
authors.forename: Dafang
authors.initials: D
authors.lastname: Huang
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Fuping
authors.initials: F
authors.lastname: Song
mesh_terms: D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000302:isolation & purification*; D003001:Cloning, Molecular / Q000379:methods; D004731:Endotoxins / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics; D006460:Hemolysin Proteins / Q000302:isolation & purification*; D007306:Insecticides / Q000302:isolation & purification*; D009036:Moths; D010572:Pest Control, Biological
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007306:Insecticides; C030956:insecticidal crystal protein, Bacillus Thuringiensis
keywords: Bacillus thuringiensis; Cry1Ac-resistance; Cry2Ah1; Helicoverpa armigera