title: [Cloning, expression and purification of allergen parvalbumin from Aristichthys nobilis and its allergic activity].
abstract: OBJECTIVE
  To clone, express and identify the parvalbumin gene from Aristichthys nobilis, and investigate its allergenicity.
  
  
  METHODS
  The parvalbumin gene was amplified by RT-PCR and cloned into PMD18-T for sequencing and analysis. Then the target gene was subcloned into pET-28a (+) for expression in E. coli BL21 (DE3) by IPTG induction. The recombinant protein was purified by metal (Ni2+) chelating affinity chromatography. Its allergenicity was examined by Western-blotting assay.
  
  
  RESULTS
  The length of gene (Accession No. FJ013047) was 330 bp, coding 109 amino acids. The E. coli strain could express a recombinant protein with a molecular weight of 11 537 Da. The recombinant allergen was identified as its affinity to specific IgE antibodies from the allergic patient sera by Western-blotting.
  
  
  CONCLUSIONS
  The parvalbumin gene from Aristichthys nobilis is successfully cloned and expressed in this study, and the recombinant protein possesses good IgE-binding capacity.
authors: Institute of Allergy and Immunology, Shenzhen University, Shenzhen 518060, China. haitun107@gmail.com
authors: Haiyan
authors: H
authors: Wang
authors: Li
authors: L
authors: Li
authors: Zhigang
authors: Z
authors: Liu
authors: Yulan
authors: Y
authors: Wu
authors: Jiajie
authors: J
authors: Chen
mesh_terms: D000485:Allergens / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029941:Fish Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D005399:Fishes; D005822:Genetic Vectors / Q000235:genetics; D007073:Immunoglobulin E / Q000276:immunology; D010320:Parvalbumins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000485:Allergens; D029941:Fish Proteins; D010320:Parvalbumins; D011994:Recombinant Proteins; D007073:Immunoglobulin E