title: Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae.
abstract: Crude extracts of house dust mites are used clinically for diagnosis and immunotherapy of allergic diseases, including bronchial asthma, perennial rhinitis, and atopic dermatitis. However, crude extracts are complexes with non-allergenic antigens and lack effective concentrations of important allergens, resulting in several side effects. Dermatophagoides farinae (Hughes; Acari: Pyroglyphidae) is one of the predominant sources of dust mite allergens, which has more than 30 groups of allergen. The cDNA coding for the group 5 allergen of D. farinae from China was cloned, sequenced and expressed. According to alignment using the VECTOR NTI 9.0 software, there were eight mismatched nucleotides in five cDNA clones resulting in seven incompatible amino acid residues, suggesting that the Der f 5 allergen might have sequence polymorphism. Bioinformatics analysis revealed that the matured Der f 5 allergen has a molecular mass of 13604.03 Da, a theoretical pI of 5.43 and is probably hydrophobic and cytoplasmic. Similarities in amino acid sequences between Der f 5 and allergens of other domestic mite species, viz. Der p 5, Blo t 5, Sui m 5, and Lep d 5, were 79, 48, 53, and 37%, respectively. Phylogenetic analysis indicated that Der f 5 and Der p 5 clustered together. Blo t 5 and Ale o 5 also clustered together, although Blomia tropicalis and Aleuroglyphus ovatus belong to different mite families, viz. Echimyopodidae and Acaridae, respectively.
authors.affiliation: Department of Laboratory Medicine, Yancheng Health Vocational and Technical College, Jiangsu Yancheng, China. ybcui1975@hotmail.com
authors.forename: Yubao
authors.initials: Y
authors.lastname: Cui
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhou
authors.forename: Guifang
authors.initials: G
authors.lastname: Ma
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.forename: Yungang
authors.initials: Y
authors.lastname: Wang
authors.forename: Weihong
authors.initials: W
authors.lastname: Shi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D039741:Antigens, Dermatophagoides / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D060829:Arthropod Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D002681:China; D003001:Cloning, Molecular; D019295:Computational Biology; D018076:DNA, Complementary; D040001:Dermatophagoides farinae / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D010802:Phylogeny; D010957:Plasmids; D017422:Sequence Analysis, DNA
chemical_list: D039741:Antigens, Dermatophagoides; D060829:Arthropod Proteins; D018076:DNA, Complementary; C451239:Dermatophagoides pteronyssinus antigen p 5