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  • Title: Cloning, expression, and analysis of the group 2 allergen from Dermatophagoides farinae from China.
    Author: Yu-bao C, Zhou Y, Weihong S, Guifang M, Yang L, Yungang W.
    Journal: An Acad Bras Cienc; 2010 Dec; 82(4):941-51. PubMed ID: 21152769.
    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.
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