Abstract：We identified new Dermatophagoides farinae allergens by DNA sequencing, cloning and prokaryotic expression of Heat shock protein 60 (Hsp60) from Dermatophagoides farinae. With degenerate primer polymerase chain reaction (PCR) to amplify the Dermatophagoides farinae cDNA, the cDNA fragments of the Hsp60 were abundantly obtained. The cDNA sequence and amino acid sequence encoded by it were detected and analysed. Meanwhile, the Hsp60 gene fragment was cloned into the pet-28a vector and transformed into E. coli for prokaryotic expression of the Hsp60. In this study, the cDNA sequence of Dermatophagoides farinae Hsp60 successfully was determined first, and the Hsp60 expressed in E. coli BL21. Results of this research laid a foundation for further analysis and study of Dermatophagoides farinae Hsp60.
[1]The International Study of Asthma and Allergies in Childhood (ISAAC) steering Committee. Worldwide variations in the prevalence of asthma symptoms: the International Study of Asthma and Allergies in Childhood (ISAAC) [R]. Eur Respir J, 1998, 12: 315-335. [2]Franzese CB, Burkhalter NW. The patient with allergies[J]. Med Clin North Am, 2010, 94(5): 891-902. [3]Maziak W, Behrens T, Brasky TM, et al. Are asthma and allergies in children and adolescents increasing? Result from ISAAC phase I and phaseⅢ surveys in Munster, Germany[J]. Allergy, 2003, 58(7): 572-579. [4]Aas K, Aberg N, Achert C, et al. European allergy white paper Allergic disease as a public health problem in Europe[R]. Braine-1’ Alleud: The UCB Institute of allergy, 1997: 116. [5]International Workshop Report. Dust mite allergens and asthma-a worldwide problem[R]. J Allergy Clin Immunol, 1989, 83: 416-427. [6]Arlian LG. Mites are ubiquitous: are mite allergens, too?[J]. Ann Allergy Asthma Immunol, 2000, 85(3): 161-163. [7]Custovic A, Simpson A, Woodcock A. Importance of indoor allergens in the induction of allergy and elicitation of allergic disease[J]. Allergy, 1998, 53(48 Suppl): 115-120. [8]Arlian LG, Platts-Mills TA. The biology of dust mites and the remediation of mite allergens in allergic disease[J]. Allergy Clin Immunol, 2001, 107(Suppl 3): S406-413. [9]Voorhorst R, Spieksma FTM, Vanekamp MJ, et al. The house dust mite (Dermatophagoides pteronyssinus) and the allergens it produces. Identity with the house dust allerge[J]. J Allergy, 1967, 39: 325-328. [10]Bessot JC, Pauli G, Mite allergens: an overview[J]. Eur Ann Allergy Clin Immunol, 2011, 43(5): 141-156. [11]Bessot JC, Pauli G. House dust mites allergens[J]. Rev Mal Respir, 2011, 28(4): 475-495. [12]Chen DX, Zhou PK, Ran PX, et al. Screening and identification of dominant allergens of Dermatophagoides pteronyssinus and Dermatophagoides farina[J]. China Trop Med,2009, 9(2): 232-234. (in Chinese) 陈代雄,周平坤,冉丕鑫,等.屋尘螨和粉尘螨主要变应原的筛选和分析[J].中国热带医学,2009,9(2):232-234. [13]Sambrook J, Russell DW. Molecular cloning: A laboratory manual [M]. 3rd ed, Beijing: Science Press, 2002: 1228-1255, 1723-1726. (in Chinese) 黄培堂,译.分子克隆实验指南[M].3版.北京:科学出版社,2002:1228-1255. [14]Puerta L, Caraballo L, Fernandez-Caldas E, et al. Nucleotide sequence analysis of a complementary DNA coding for a Blomia tropicalis allergen[J]. J Allergy Clin Immunol, 1996, 98(5 Pt 1): 932-937. [15]Young RA. Stress proteins and immunology[J]. Annual Rev Immunol, 1990, 8: 401-420. [16]Karamura HY, Hancock K, Rodrigues V, et al. Schistosoma mansoni heat shock protein 70 elicits an early humoral immune response in S. mansoni infected baboons[J]. Mem Inst Oswaldo Cruz, 2002, 37(5): 711-716. [17]Zhang M, Hisaeda H, Kano S, et al. Antibodies specific for heat shock proteins in human and murine malaria[J]. Microbes Infect, 2001, 3(5): 363-367. [18]Aki T, Fujikawa A, Wada T, et al. Cloning and expression of cDNA coding for a new allergen from the house dust mite, Dermatophagoides farinae: homology with human heat shock cognate proteins in the heat shock protein 70 family[J]. J Biochem, 1994, 115(3): 435-440.
2015, Vol. 31 
