Abstract: Objective A chimeric gene library of the dust mites group 2 allergen, Der f2 and Der p2, was constructed by using DNA shuffling method and analyzed by bioinformatics. Method Der f2 and Der p2 genes were amplified by polymerase chain reaction (PCR), respectively; the PCR products were mixed equally (v/v) and digested with DNase I. DNA fragments of 50bp-100bp were purified. DNA shuffling was performed through three rounds of primerless PCR and additional PCR with Der f2 specific primers. A single product of correct size was inserted into pUCm-T vector and transformed into E. coli DH-5α. Sixty clones were picked out randomly, sequenced and analyzed by multiple sequence alignment using DNAMAN software. T helper cell epitopes were predicted by NetMHCII 2.2 Server. Results Ten recombinants were obtained. The amino acid sequences deduced from these recombinants were predicted to not only share the common Th cell epitopes with Der f2 and Der p2 but also possess new epitopes. Conclusion A chimeric gene library of Der f2 and Der p2 was successfully established, which could provide the basis for selection of vaccine with high immunogenicity and low allergenicity for asthma treatment in future.

Key words: allergens, chimeric gene, DNA shuffling