title: Constructing Bacillus thuringiensis strain that co-expresses Cry2Aa and chitinase.
abstract: A triple recombineering technique was used with plasmid pHT315 to produce pHTEC, a construct carrying chitinase and cry2Aa genes from Bacillus thuringiensis subsp. kurstaki 4.0718. Transformation of wild-type B. thuringiensis strain HD73 and the acrystalliferous strain Cry-B with pHTEC resulted in the recovery of recombinant strains that expressed Cry2Aa as cubic crystals in the cell pellet and soluble chitinase protein. The toxicity of HD73 (pHTEC) against Helicoverpa armigera larvae increased sevenfold when compared with HD73 (pHT315) harboring pHT315 vector. The triple recombineering protocol was optimized by comparing recombination efficacy mediated by RecE/RecT and Redα/Redβ and by using single-strand DNA as substrate.
authors.affiliation: Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, College of Life Science, Hunan Normal University, Changsha, China. hsbalp@163.com
authors.forename: Shengbiao
authors.initials: S
authors.lastname: Hu
authors.forename: Xu
authors.initials: X
authors.lastname: Zhang
authors.forename: Yusheng
authors.initials: Y
authors.lastname: Li
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Ding
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Hu
authors.forename: Qi
authors.initials: Q
authors.lastname: Yang
authors.forename: Liqiu
authors.initials: L
authors.lastname: Xia
mesh_terms: D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics* / Q000378:metabolism*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001681:Biological Assay; D002688:Chitinases / Q000235:genetics / Q000378:metabolism*; D004731:Endotoxins / Q000235:genetics / Q000378:metabolism*; D006460:Hemolysin Proteins / Q000235:genetics / Q000378:metabolism*; D007814:Larva / Q000382:microbiology / Q000502:physiology; D007915:Lepidoptera / Q000382:microbiology* / Q000502:physiology*; D010957:Plasmids; D016019:Survival Analysis
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D002688:Chitinases