title: [Expression of goat IL-18 mature protein in insect/baculovirus and determination of bioactivity of the recombinant protein].
abstract: OBJECTIVE
  To express goat IL-18 in insect/baculovirus and detect the bioactivity of the recombinant protein.
  
  
  METHODS
  The mature goat interleukin-18(gIL-18) gene was cloned into the baculovirus transfer vector pFastBac Dual, and then the resulting eukaryotic expression plasmid pFastBac Dual-gIL18 was transformed into DH10Bac, followed by the identification of Bacmid-gIL18 recombinat plosmid by three antibiotics and blue-white patch. Finally, the recombinant bacmid was transfected into sf9 insect cells by Cellfectin and the transfected cells were harvested at different times. Then the expressed protein was identified by SDS-PAGE, Western blot and bioactivity assay.
  
  
  RESULTS
  The recombinant protein recognized and bound to its specific antibody. Bioactivity assay showed that the recombinant protein stimulated the proliferation of lymphocytes and induced IFN-γproduction in spleen lymphocytes.
  
  
  CONCLUSIONS
  The mature gIL-18 protein has been expressed successfully in insect/baculovirus expression system, and have good immunogenicity and bioactivity. The study paves a way for application of gIL-18 as an immunomodulator or immune adjuvant.
authors.affiliation: Shandong Engineering Research Center for Meat Food Quality Safety, College of Food Science and Engineering, Qingdao Agricultural University, Qingdao 266109, China.
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Wang
authors.forename: Xi-Hui
authors.initials: XH
authors.lastname: Wang
authors.forename: Zhong-Ling
authors.initials: ZL
authors.lastname: Fan
authors.forename: Jin-Long
authors.initials: JL
authors.lastname: Chen
authors.forename: Bing-Lei
authors.initials: BL
authors.lastname: Cao
authors.forename: Na
authors.initials: N
authors.lastname: Kong
authors.forename: Jing-Dong
authors.initials: JD
authors.lastname: Hu
authors.forename: Hong-Kun
authors.initials: HK
authors.lastname: Zhao
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006041:Goats; D007313:Insecta / Q000166:cytology; D007371:Interferon-gamma / Q000378:metabolism; D020382:Interleukin-18 / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D018116:Vesiculovirus / Q000187:drug effects
chemical_list: D020382:Interleukin-18; D011994:Recombinant Proteins; D007371:Interferon-gamma