DNA shuffling and bioinformatics analysis of the dust mites group 2 allergen genes,Der f2 and Der p2
Ma Yu-cheng, Zhu Tao, Jiang Yu-xin
2012
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0
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Journal
Basic & Clinical Medicine
Semantic Scholar
Key Takeaway Key Takeaway: A chimeric gene library of dust mite allergens Der f2 and Der p2 has been successfully established, potentially aiding in the development of high immunogenic and low allergenic vaccines for asthma treatment.
Abstract
Objective To construct and analyze a chimeric gene library of the dust mites group 2 allergen,Der f2 and Der p2 gene by DNA shuffling method and bioinformatics respectively. Methods Der f2 and Der p2 genes were amplified by polymerase chain reaction( PCR) , respectively; the PCR products were mixed equally( v / v) and di- gested with DNase Ⅰ . Fragments from 50 to 100 bp were purified,reassembled by PCR without primers for 3 times continously,and amplified with specific Der f2 primers at the last. The reassembled product of original size was in- serted into pUCm-T vector and transformed into E. coli DH-5 α . Sixty clones were randomly picked out,sequenced and aligned using DNAMAN software. The epitopes of T helper ( Th) cell and B cell were deduced by NetMHCII 2. 2 Server. Results The amino acid sequences deduced from 10 recombinants not only increased new Th cell epitopes but also decreased B cell epitopes as compared with original genes. Conclusions A chimeric gene library from Der f2 and Der p2 was successfully established which may provide the basis for selection of vaccine with high immunogenicity and low allergenicity applyed in asthma treatment.