title: [Cloning, expression and identification of Der f7 gene from Dermatophagoides farinae and its immunological characteristics].
abstract: OBJECTIVE
  To clone and express Der f7 gene of Dermatophagoides farinae, and identify its immunogenicity.
  
  
  METHODS
  Total RNA was extracted from D. farinae mites. A reference sequence (Accession No. AY283292) was used to design specific primers. The Der f7 gene fragment was amplified by RT-PCR, and cloned into pET-32a vector. The recombinant plasmid was transformed into E. coli BL21 (DE3) and induced with IPTG for protein expression. The recombinant protein was purified by Ni2+ chelating affinity chromatography and analyzed by SDS-PAGE and Western blotting.
  
  
  RESULTS
  The Der f7 gene fragment was about 650 bp, and shared 99% homology with the published one (Accession No. FJ436108). SDS-PAGE result showed its relative molecular weight (M(r)) of 23 000. The recombinant protein showed appropriate combination ability with IgE in sera of mite allergic patients.
  
  
  CONCLUSIONS
  Der f 7 gene has been expressed in prokaryotic expression system and shows allergenicity.
authors.forename: Man-Yin
authors.initials: MY
authors.lastname: Zheng
authors.forename: Yu-Lan
authors.initials: YL
authors.lastname: Wu
authors.forename: Hao
authors.initials: H
authors.lastname: Yan
authors.forename: Kun-Mei
authors.initials: KM
authors.lastname: Ji
authors.forename: Zhi-Gang
authors.initials: ZG
authors.lastname: Liu
mesh_terms: D000485:Allergens / Q000235:genetics / Q000276:immunology; D000818:Animals; D039741:Antigens, Dermatophagoides / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D060829:Arthropod Proteins / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D003001:Cloning, Molecular; D040001:Dermatophagoides farinae / Q000235:genetics* / Q000276:immunology; D015870:Gene Expression; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000485:Allergens; D039741:Antigens, Dermatophagoides; D060829:Arthropod Proteins; C451161:Dermatophagoides farinae antigen f 7; D011994:Recombinant Proteins