title: Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen.
abstract: Domestic mite species like Dermatophagoides farinae induce allergies in people worldwide. Here, the cDNA coding for group 21 allergen of Dermatophagoides farinae (Hughes; Acari: Pyroglyphidae) from China was cloned, sequenced, and expressed in E. coli to aid in the development of diagnostic and treatment options for domestic mite hypersensitivity. First, the Der f 21 cDNA fragment was synthesized by RT-PCR; the confirmed full-length sequence comprised 411 nucleotides. The cDNA was ligated to the vector pCold-TF to construct an expression plasmid, pCold-TF-Der f 21. pCold-Tf-Der f 21 was transformed into E. coli BL21 cells, and its expression was induced by IPTG treatment. SDS-PAGE showed a specific band at the predicted molecular weight of Der f 21, demonstrating its successful expression. The recombinant fusion protein was obtained and its structure and molecular weight were confirmed by MALDI-TOF/TOF. Bioinformatics analysis revealed that the protein contained a signal peptide of 17 amino acids. The molecular weight of the mature Der f 21 allergen was approximately 14.16 kDa with a theoretical pI of 4.87. Its predicted secondary structure comprises a-helix (84.03%), extension chain (1.68%), and random coil (14.29%). The successful cloning of Der f 21 and a basic bioinformatics analysis of the protein provide a foundation for further study of this allergen in diagnosis and treatment of domestic mite hypersensitivity.
authors: Department of Laboratory Medicine, Shanghai Punan Hospital Pudong New District, Shanghai 200125, P. R. China ; Department of Laboratory Medicine, Yancheng Health Vocational & Technical College Jiangsu Yancheng 224006, P. R. China.
authors: Yubao
authors: Y
authors: Cui
authors: Department of Respiratory, The First People's Hospital of Yancheng City Jiangsu Yancheng 224005, P. R. China.
authors: Yongqian
authors: Y
authors: Jiang
authors: Department of Respiratory, The First People's Hospital of Yancheng City Jiangsu Yancheng 224005, P. R. China.
authors: Youlin
authors: Y
authors: Ji
authors: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College Jiangsu Yancheng 224006, P. R. China.
authors: Ying
authors: Y
authors: Zhou
authors: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College Jiangsu Yancheng 224006, P. R. China.
authors: Lili
authors: L
authors: Yu
authors: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College Jiangsu Yancheng 224006, P. R. China.
authors: Nan
authors: N
authors: Wang
authors: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College Jiangsu Yancheng 224006, P. R. China.
authors: Li
authors: L
authors: Yang
authors: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College Jiangsu Yancheng 224006, P. R. China.
authors: Chengbo
authors: C
authors: Zhang
keywords: Der f 21; Dermatophagoides farinae; domestic mite; molecular cloning; recombinant allergen
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