title: Improved insecticidal toxicity by fusing Cry1Ac of Bacillus thuringiensis with Av3 of Anemonia viridis.
abstract: Av3, a neurotoxin of Anemonia viridis, is toxic to crustaceans and cockroaches but inactive in mammals. In the present study, Av3 was expressed in Escherichia coli Origami B (DE3) and purified by reversed-phase liquid chromatography. The purified Av3 was injected into the hemocoel of Helicoverpa armigera, rendering the worm paralyzed. Then, Av3 was expressed alone or fusion expressed with the Cry1Ac in acrystalliferous strain Cry(-)B of Bacillus thuringiensis. The shape of Cry1Ac was changed by fusion with Av3. The expressed fusion protein, Cry1AcAv3, formed irregular rhombus- or crescent-shaped crystalline inclusions, which is quite different from the shape of original Cry1Ac crystals. The toxicity of Cry1Ac was improved by fused expression. Compared with original Cry1Ac expressed in Cry(-)B, the oral toxicity of Cry1AcAv3 to H. armigera was elevated about 2.6-fold. No toxicity was detected when Av3 was expressed in Cry(-)B alone. The present study confirmed that marine toxins could be used in bio-control and implied that fused expression with other insecticidal proteins could be an efficient way for their application.
authors.affiliation: Hunan Provincial Key Laboratory for Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology and College of Life Science, Hunan Normal University, Changsha, 410081, People's Republic of China.
authors.forename: Fu
authors.initials: F
authors.lastname: Yan
authors.forename: Xing
authors.initials: X
authors.lastname: Cheng
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Ding
authors.forename: Ting
authors.initials: T
authors.lastname: Yao
authors.forename: Hanna
authors.initials: H
authors.lastname: Chen
authors.forename: Wenping
authors.initials: W
authors.lastname: Li
authors.forename: Shengbiao
authors.initials: S
authors.lastname: Hu
authors.forename: Ziquan
authors.initials: Z
authors.lastname: Yu
authors.forename: Yunjun
authors.initials: Y
authors.lastname: Sun
authors.forename: Youming
authors.initials: Y
authors.lastname: Zhang
authors.forename: Liqiu
authors.initials: L
authors.lastname: Xia
mesh_terms: D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics / Q000633:toxicity*; D001681:Biological Assay; D004731:Endotoxins / Q000235:genetics / Q000633:toxicity*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006460:Hemolysin Proteins / Q000235:genetics / Q000633:toxicity*; D007306:Insecticides / Q000633:toxicity*; D007915:Lepidoptera / Q000187:drug effects; D009498:Neurotoxins / Q000235:genetics / Q000633:toxicity*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000633:toxicity; D012615:Sea Anemones / Q000235:genetics*; D016019:Survival Analysis
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007306:Insecticides; D009498:Neurotoxins; D011993:Recombinant Fusion Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis