title: [The gene expression of the protein SLAWD, mediating the toxic effect of destruxin A on Spodoptera litura larvae, in procaryotic cells: purification and characterization].
abstract: Spodoptera litura is one of the most destructive phytophagous pest infesting cotton, vegetable, oilseed and ber crops around the world. Dextruxin A (DA), is a one of a kind microbial insecticide, which has potent toxins with bioactivity against S. litura larvae. An abnormal wing disc (AWD) protein was identified as a DA toxic effect protein in S. litura SL-1 cells. To better understand the role of the AWD gene of S. litura (SLAWD) it was purified and characterized. The entire coding region of the SLAWD gene was cloned into a pET-32a(+) expression vector and transformed into competent Escherichia coli BL21 (DE3) cells. SDS-PAGE and western blotting analysis and western blotting showed that the best induction conditions were 1 mmol mL(-1) isopropyl-β-D-thiogalactopyranoside (IPTG) for 6 h at 37°C; the molecular weight of the fusion protein was 35.0 kDa. The production of polyclonal antibodies and an enzyme-linked immunosorbent assay (ELISA) showed that the titer of antiserum was 1:25,600; western blotting analysis showed that the recombinant SLAWD was recognized by the anti-SLAWD polyclonal antibody. AWD is a key protein involved in wing development in insects. These tools will assist in the further characterization of SLAWD and studies on the mechanism of action of destruxin A.
authors.forename: X
authors.initials: X
authors.lastname: Meng
authors.forename: L M
authors.initials: LM
authors.lastname: Li
authors.forename: G
authors.initials: G
authors.lastname: Gao
authors.forename: F L
authors.initials: FL
authors.lastname: Jin
authors.forename: S X
authors.initials: SX
authors.lastname: Ren
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000302:isolation & purification; D003001:Cloning, Molecular; D047630:Depsipeptides / Q000633:toxicity; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D019476:Insect Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D007814:Larva / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology / Q000254:growth & development; D009183:Mycotoxins / Q000633:toxicity; D009701:Nucleoside-Diphosphate Kinase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D016366:Open Reading Frames; D010957:Plasmids / Q000737:chemistry*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D018411:Spodoptera / Q000737:chemistry* / Q000187:drug effects / Q000201:enzymology / Q000254:growth & development; D014921:Wings, Animal / Q000737:chemistry / Q000201:enzymology / Q000254:growth & development
chemical_list: D000906:Antibodies; D047630:Depsipeptides; D019476:Insect Proteins; D009183:Mycotoxins; D011993:Recombinant Fusion Proteins; C040423:destruxin A; D009701:Nucleoside-Diphosphate Kinase