title: Cloning, expression, and analysis of the group 2 allergen from Dermatophagoides farinae from China.
abstract: To obtain the recombinant group 2 allergen product of Dermatophagoides farinae (Der f 2), the Der f 2 gene was synthesized by RT-PCR. The full-length cDNA comprised 441 nucleotides and was 99.3% identical to the reference sequence (GenBank AB195580). The cDNA was bound to vector pET28a to construct plasmid pET28a(+)-Der f 2, which was transformed into E. coli BL21 and induced by IPTG. SDS-PAGE showed a specific band of about 14kDa in the hole cell lysate. As estimated by chromatography, about 3.86 g of the recombinant product as obtained, which conjugated with serum IgE from asthmatic children. The protein had a signal peptide of 17 amino acids. Its secondary structure comprised an alpha helix (19.86%), an extended strand (30.82%), and a random coil (49.32%). The subcellular localization of this allergen was predicted to be at mitochondria. Furthermore, its function was shown to be associated with an MD-2-related lipid-recognition (ML) domain. The results of this study provide a solid foundation for large-scale production of the allergen for clinical diagnosis and treatment of allergic disorders.
authors.affiliation: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College, Jiangsu Yancheng, PR China. ybcui1975@hotmail.com
authors.forename: Cui
authors.initials: C
authors.lastname: Yu-bao
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhou
authors.forename: Shi
authors.initials: S
authors.lastname: Weihong
authors.forename: Ma
authors.initials: M
authors.lastname: Guifang
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.forename: Wang
authors.initials: W
authors.lastname: Yungang
mesh_terms: D000485:Allergens / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D039741:Antigens, Dermatophagoides / Q000737:chemistry / Q000235:genetics*; D015153:Blotting, Western; D002675:Child, Preschool; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D040001:Dermatophagoides farinae / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
chemical_list: D000485:Allergens; D039741:Antigens, Dermatophagoides; D018076:DNA, Complementary