粉尘螨2类变应原基因的克隆和序列分析  

Cloning and Sequencing of Dermatophagoides Farinae Major Antigen Der f 2

作　　者：张理涛[1] 纪岩文[2] 张铁楠[2] 张玉环[1] 

机构地区：[1]天津市长征医院皮肤变态反应科,300120 [2]天津医科大学总医院皮肤科

出　　处：《天津医药》2008年第9期666-668,共3页Tianjin Medical Journal

摘　　要：目的:克隆中国大陆地区粉尘螨2类变应原(Derf2)的cDNA。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Derf2基因,克隆至pMD18-TSimple平滑载体中测序。结果:筛选出2株长度为436bp的Derf2cDNA片段,核苷酸序列1株与Genbank上的Derf2序列(D10448)碱基有6处不同,同源性为98.6%;另1株有2处不同,同源性为99.7%。结论:粉尘螨2类变应原的基因序列具有多态性,中国大陆地区Derf2的cDNA克隆片段与Genbank上公布的Derf2序列高度同源。Objective： To clone and analyze the cDNA of major allergen of Chinese Dermatophagoides farinae（Der f 2）. Methods： The total RNA of Dermatophagoides farinae was extracted by Trizol. The cDNA of Der f 2 was amplified with RT-PCR,and then they were cloned into the vector pMD18-T simple and sequenced. Results： Two segments with 436 bases were cloned. Compared to the sequence homology of Der f 2（D10448）published on GenBank,one was 98.6% at the nucleotide level with 6 different bases,the other was 99.7% with 2 different bases. Conclusion： The gene sequences of Der f 2 of the dust mite showed polymorphism,which indicated that its sequence homology was high as compared with the published one in GenBank.

关 键 词：粉螨科 变应原 DNA 互补 序列分析 DNA 克隆 分子 逆转录聚合酶链反应 

分 类 号：R373.21[医药卫生—病原生物学] R394[医药卫生—基础医学]