Cloning and analysis of the antigen-coding gene Der f 1 from house dust mite Dermatophagoides farinae in Escherichia coli

Abstract

Der f 1, an allergen from Dermatophagoides farinae (D. farinae), is one of the major causes of house dust mite (HDM) allergy. The aim of this study was to clone the gene encoding the Der f 1 allergen from D. farinae collected in Vietnam, providing a basis for the development of recombinant Der f 1 protein. The amplified product containing the full-length coding sequence of Der f 1 (obtained through a three-step PCR using cDNA synthesised from the RNA of D. farinae) was cloned into the pGEM-T vector and transformed into Escherichia coli DH5α. The recombinant plasmids were screened by colony PCR and sequenced using the Sanger method. The sequence of Der f 1 and its deduced amino acid had 99% similarities to the reference nucleotide and amino acid sequences in GenBank, with the accession number of AB034946.1 and ABL84751.1, respectively. The Der f 1 gene sequence from D. farinae collected in Vietnam has been deposited in GenBank under the accession number PP417680.1. The results of the study provide essential foundational data for further research on recombinant Der f 1 protein, supporting the diagnosis and treatment of house dust mite allergy through specific immunotherapy in Vietnam.