CAJ | 학술논문

目的：克隆粉尘螨过敏原Der f 11基因，表达、纯化该蛋白，并鉴定其免疫活性。方法：人工合成粉尘螨第11组过敏原Der f 11基因，将其连接至pET-32a表达载体，用异丙基-β-D-硫代半乳糖苷（IPTG）诱导表达后，通过Ni＋亲和层析纯化重组过敏原Der f 11。以粉尘螨过敏患者血清作为一抗，经Western blot方法分析Der f 11的免疫学特性。结果：获得高纯度的重组Der f 11蛋白，SDS-PAGE结果显示表达产物pET-32a（＋）-Der f 11分子质量约为118 ku。重组过敏原Der f 11检测15份尘螨过敏性患者血清中特异性IgE，阳性率为20％。结论：获得的Der f 11重组蛋白具有与天然蛋白相似的免疫学活性，为标准化抗原的临床特异性诊断和治疗及进一步的实验研究奠定基础。
목적：극륭분진만과민원Der f 11기인，표체、순화해단백，병감정기면역활성。방법：인공합성분진만제11조과민원Der f 11기인，장기련접지pET-32a표체재체，용이병기-β-D-류대반유당감（IPTG）유도표체후，통과Ni＋친화층석순화중조과민원Der f 11。이분진만과민환자혈청작위일항，경Western blot방법분석Der f 11적면역학특성。결과：획득고순도적중조Der f 11단백，SDS-PAGE결과현시표체산물pET-32a（＋）-Der f 11분자질량약위118 ku。중조과민원Der f 11검측15빈진만과민성환자혈청중특이성IgE，양성솔위20％。결론：획득적Der f 11중조단백구유여천연단백상사적면역학활성，위표준화항원적림상특이성진단화치료급진일보적실험연구전정기출。
Objective: To clone , express and purify of Dermatophagoides farinae ( Der f 11 ) , and then test its immunogenicity.Methods:The gene coding for Der f 11 was synthesized ,and was then linked with the pET-32a vector.The expression plasmid pET32a(+)-Der f 11 was induced by IPTG.After purification of recombinant allergens Der 11 proteins through the Ni +affinity chromatography ,immunological allergic patients serum as the Primary antibody.Results: We obtain high purity recombinant Der f 11 protein.The results of SDS-PAGE show that the expression product is about 118 KD.Recombinant allergen Der f 11 test 15 dust mites allergic patients serum specific IgE , positive rate was 20%.Conclusion: Recombinant allergen Der f 11 obtained has the similar immunologic activity to natural Der f 11 protein.It can lay the foundation for the specific diagnosis ,treatment and further experimental studies of the dust mite allergy disease.