基础医学与临床2012，Vol.32Issue(6)：634-638,5.
尘螨Ⅱ类变应原Der f2和Der p2的DNA改组及生物信息学分析
DNA shuffling and bioinformatics analysis of the dust mites group 2 allergen genes, Der f2 and Der p2
摘要
Abstract
Objective To construct and analyze a chimeric gene library of the dust mites group 2 allergen, Derf2 and Der p2 gene by DNA shuffling method and bioinformatics respectively. Methods Der ft and Der p2 genes were amplified by polymerase chain reaction (PCR), respectively; the PCR products were mixed equally (v/v) and digested 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 ft. primers at the last. The reassembled product of original size was inserted into pUCm-T vector and transformed into E. coli DH-5α. 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.关键词
变应原/嵌合基因/DNA改组Key words
allergens/ chimeric gene/ DNA shuffling分类
医药卫生引用本文复制引用
马玉成,朱涛,姜玉新,李朝品..尘螨Ⅱ类变应原Der f2和Der p2的DNA改组及生物信息学分析[J].基础医学与临床,2012,32(6):634-638,5.基金项目
国家自然科学基金(30872367:81172790) （30872367:81172790）
安徽省自然科学基金(070413088) （070413088）
