关键词: 尘螨, Der p 2, 克隆, 原核表达

Abstract: Objective To clone, express and characterize Dermatophagoides pteronyssinus major allergens. Methods Based on Der p 2 nucleotide sequence published in the GenBank, we designed primers and amplified the cDNA fragment coding Der p 2 allergen from D.pteronyssinus by RT-PCR. After purification and recovery, the cDNA fragment was cloned into the pMD19-T vector. The fragment was then sequenced, subcloned into the plasmid pET28a(+), expressed in Escherichia coli BL21 and identified by Western blotting. Results The cDNA coding Derp 2 allergen of adultD.pteronyssinus was cloned, sequenced and expressed successfully. Sequence analysis showed the gene homology with Der p 2 reported in GenBank was 99.54%, which probably encode a protein with 130 amino acids and its molecular weight was 14 348.5. Conclusion The cDNA coding Derp 2 allergen ofD.pteronyssinus were cloned and expressed successfully, which provided a foundation for the production of recombined allergenst.