Cloning, expression, and analysis of the group 2 allergen from Dermatophagoides farinae from China

Anais da Academia Brasileira de Ciências

Cui Yu-bao¹  Ying Zhou¹  Shi Weihong¹  Ma Guifang¹  Li Yang¹  Wang Yungang¹ 

[1] ,Yancheng Health Vocational & Technical College Department of Laboratory Medicine Jiangsu Yancheng,P.R. China

关键词: Dermatophagoides farinae;    house-dust-mite allergy;    mite allergens;    Der f 2;    allergen engineering;    immunotherapy;    bioinformatics;    Dermatophagoides farinae;    alergia a ácaro de poeira doméstica;    alergenos acarianos;    Der f 2;    alergeno recombinante;    imunoterapia;    bioinformática;   

DOI  :  10.1590/S0001-37652010000400017

来源: SciELO

【 摘 要 】

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. s estiated by chroatography, about 3.86 g of the recobinant 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 treatent of allergic disorders.

【 授权许可】

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