title: Expression and characterization of a recombinant Cry1Ac crystal protein fused with an insect-specific neurotoxin ω-ACTX-Hv1a in Bacillus thuringiensis.
abstract: In order to assess possible enhancement of biopesticide activity, the fusion gene of crystal protein gene cry1Ac with the insect-specific neurotoxin ω-ACTX-Hv1a gene and egfp was expressed in Bacillus thuringiensis acrystalliferous strain Cry-B under the control of the native gene expression system. The fusion recombinant Cry-B(1Ac-ACTX-EGFP) generally produced two or three small crystal-like inclusion bodies in each cell and the GFP signal could be clearly observed. A 166 kDa full-length fusion protein was identified by immunoblot analysis. Virulence of the fusion inclusions was at least fivefold higher toward larvae of Spodoptera exigua. These results demonstrated that a foreign protein could be expressed and accumulate as parasporal inclusions in B. thuringiensis by C-terminal fusion with the native endotoxin while retaining partial insecticidal activity.
authors.affiliation: College of Life Science, Hunan Normal University, State Key Laboratory of Breeding Base of Microbial Molecular Biology, Changsha 410081, PR China.
authors.forename: W P
authors.initials: WP
authors.lastname: Li
authors.forename: L Q
authors.initials: LQ
authors.lastname: Xia
authors.forename: X Z
authors.initials: XZ
authors.lastname: Ding
authors.forename: Y
authors.initials: Y
authors.lastname: Lv
authors.forename: Y S
authors.initials: YS
authors.lastname: Luo
authors.forename: S B
authors.initials: SB
authors.lastname: Hu
authors.forename: J
authors.initials: J
authors.lastname: Yin
authors.forename: F
authors.initials: F
authors.lastname: Yan
mesh_terms: D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics / Q000378:metabolism*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004731:Endotoxins / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006460:Hemolysin Proteins / Q000235:genetics* / Q000378:metabolism; D002479:Inclusion Bodies; D007306:Insecticides / Q000494:pharmacology*; D007814:Larva / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D013111:Spider Venoms / Q000235:genetics* / Q000378:metabolism; D018411:Spodoptera / Q000187:drug effects
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007306:Insecticides; D011993:Recombinant Fusion Proteins; D013111:Spider Venoms; C030956:insecticidal crystal protein, Bacillus Thuringiensis; C107148:omega-atracotoxin-HV1; D049452:Green Fluorescent Proteins