title: [Expression and characterization of the dermonecrotic toxin gene of Bordetella bronchiseptica].
abstract: Dermonecrotic toxin (DNT) is identified as one of the most important virulence factor of Bordetella bronchiseptica. The complete coding sequence (4 356 bp) of the dnt gene was cloned into the prokaryotic expression vector pET-28a, and expressed in the Eschierichia coli BL21 (DE3) under IPTG (Isopropyl-beta-D-thiogalactopyranoside) induction. The recombinant His6-DNT protein showed immunological reactivity in the Western-blot analysis. The recombinant protein was purified from crude lysates of BL21 harboring pET-DNT with the purity of 93.2%. His6-DNT showed the dermonecrotic effects in the infant mouse assay. However, rabbit anti-serum against recombinant DNT protein could neutralize the dermonecrotic effects of native DNT to the infant mice in vivo. These findings suggest that the recombinant DNT protein retained the characteristics and immunogenicity of native DNT. Furthermore, this approach could be used to induce active immunity and serum immunoglobulin for production of a passive therapeutic reagent. In this study, we have shown that the recombinant His6-DNT protein retained the characteristics of native DNT of B. bronchiseptica, which built a good foundation for the further research on the structure and function of DNT.
authors.affiliation: College ofAnimal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Xue
authors.forename: Zhanqin
authors.initials: Z
authors.lastname: Zhao
authors.forename: Jie
authors.initials: J
authors.lastname: Pei
authors.forename: Chen
authors.initials: C
authors.lastname: Wang
authors.forename: Ke
authors.initials: K
authors.lastname: Ding
authors.forename: Xiangchao
authors.initials: X
authors.lastname: Cheng
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D016950:Bordetella bronchiseptica / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D009500:Neutralization Tests; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011503:Transglutaminases / Q000096:biosynthesis* / Q000235:genetics*; D010566:Virulence Factors, Bordetella / Q000096:biosynthesis* / Q000235:genetics*
chemical_list: D011993:Recombinant Fusion Proteins; D010566:Virulence Factors, Bordetella; C060969:dermonecrotic toxin, Bordetella; D011503:Transglutaminases