title: [Immuno-cross-reactivity of different antifreeze proteins from Tenebrionidae insects].
abstract: OBJECTIVE
  To prepare the antiserum against antifreeze protein, MpAFP698, from the beetle Microdera punctipennis (M. punctipennis) and determine immunological homology of different antifreeze proteins from different insects in Tenebrionidae.
  
  
  METHODS
  Two New Zealand white rabbits were immunized by DNA prime-protein boost strategy. Firstly, recombinant plasmid pcDNA3-Mpafp698 was used as DNA vaccine to inoculate two rabbits respectively for two times. Then twice boost immunization was performed using recombinant protein His-MpAFP698. ELISA and Western blotting were respectively conducted to determine the titer of MpAFP698 antibody and to detect the specificity of the antibody as well as the immunological cross-reaction of different insect antifreeze proteins with MpAFP698 antibody.
  
  
  RESULTS
  The titer of the rabbit antiserum reached 1:400 000. Western blotting showed the specific binding of the antiserum to His-MpAFP698 protein and GST-MpAFP698 protein respectively. The rabbit antiserum also presented specific binding to antifreeze protein MpAFP149 and MpAFPS77 from M.punctipennis, as well as Apafp914 and OcAFP4 from different species.
  
  
  CONCLUSIONS
  Antifreeze proteins from different beetles in desert region have the same epitope and thus have immunological cross-reaction.
authors.affiliation: College of Life Science and Technology, Xinjiang University, Urumqi, China.
authors.forename: Jieqiong
authors.initials: J
authors.lastname: Li
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Hou
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.forename: Xiaoning
authors.initials: X
authors.lastname: Liu
authors.forename: Ji
authors.initials: J
authors.lastname: Ma
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D021301:Antifreeze Proteins / Q000276:immunology*; D001517:Coleoptera; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D007106:Immune Sera / Q000276:immunology; D019476:Insect Proteins / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology*
chemical_list: D021301:Antifreeze Proteins; D007106:Immune Sera; D019476:Insect Proteins; D011993:Recombinant Fusion Proteins