粉尘螨1类变应原(Der f1)的cDNA克隆和序列分析

Cloning and Sequencing of Major Allergen Der f1 cDNA of Dermatophagoides Farinae

目的获得中国大陆地区粉尘螨1类变应原(Der f1)cDNA序列,为进一步表达重组的Der f1变应原用于尘螨过敏性疾病的诊治打下基础。方法挑取活的粉尘螨经-70℃冷冻后提取总RNA,采用RT-PCR的方法扩增Der f1片断,进行克隆、测序。结果筛选出2株长度为361bp的Der f1 cDNA片断,一株核苷酸序列与Genebank上的Der f1序列(AB034946)完全相同;另一株与Der f1序列比较有2处不同,同源性为99.7%。结论粉尘螨变应原的序列具有多态性,我们获得的中国大陆地区Der f1的cDNA克隆片断与Genebank上公布的Derf1序列高度同源。

Objective To clone and analyze the major allergen Der f1 cDNA of Chinese Dermatophagoides farinae for further diagnosis and therapy purpose of dust mite allergic diseases.Methods The total RNA of Dermatophagoides farinae was extracted by Trizol.The cDNA of Der f1 was amplified with RT-PCR,then were cloned into the vector pMD18-T Simple and sequenced.Results Two segments with 361 bases were cloned.Compared the sequence homology with the published Der f1（AB034946）on Genebank,one was 100% at the nucleotide level,the other was 99.7% with two different bases.Conclusion The gene sequences of Der f1 of the dust mite showed polymorphism,which indicated that the allergens of Dermatophagoides farinae be multiple and the therapy of allergic diseases be complicated.