title: Characterization of the chaperonin GroEL in Mycoplasma gallisepticum.
abstract: Mycoplasma gallisepticum (MG) is a common and widespread cause of chronic respiratory disease in poultry. In this study, antigenic proteins were identified from MG membrane using two-dimensional gel electrophoresis (2-DE) analysis followed by Western blot and matrix-assisted desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), including translation elongation factor Tu, dihydrolipoamide acetyltransferase (E2) component of pyruvate dehydrogenase complex, trigger factor, chaperone protein DnaK, heat shock protein GroEL and so on. Furthermore, recombinant MG GroEL protein was successfully expressed in E. coli BL21 (DE3) with pET-28a (+) vector and found to possess ATPase activity and contributed to the refolding of recombinant MG PrpC protein. Complement-dependent bactericidal assay indicated that the rabbit antisera against MG rGroEL had satisfactory bactericidal effect, which is similar to the chicken antisera induced by MG-inactivated vaccine, suggesting MG GroEL is a protective antigen, could be used as a novel vaccine candidate. This study is the first report of the biological characterization of chaperone GroEL protein in MG.
authors.affiliation: Department of Avian Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No.518, Ziyue Road, Minhang District, Shanghai, 200241, People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: Tan
authors.forename: Meirong
authors.initials: M
authors.lastname: Hu
authors.forename: Shengqing
authors.initials: S
authors.lastname: Yu
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Feng
authors.initials: F
authors.lastname: Lu
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
authors.forename: Xusheng
authors.initials: X
authors.lastname: Qiu
authors.forename: Cuiping
authors.initials: C
authors.lastname: Song
authors.forename: Yingjie
authors.initials: Y
authors.lastname: Sun
authors.forename: Chan
authors.initials: C
authors.lastname: Ding
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D000942:Antigens, Bacterial / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D018834:Chaperonin 60 / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D007106:Immune Sera / Q000494:pharmacology; D045686:Mycoplasma gallisepticum / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D058248:Serum Bactericidal Antibody Assay
chemical_list: D000942:Antigens, Bacterial; D018834:Chaperonin 60; D007106:Immune Sera; D011994:Recombinant Proteins; D000251:Adenosine Triphosphatases