表达HPCagA基因非复制型重组腺病毒的构建

目的：构建表达幽门螺杆菌CagA基因的非复制型重组腺病毒，从而为探讨表达HpCagA的重组腺病毒对哮喘TH1/TH2细胞因子的调控作用，获得防治哮喘的新方法奠定基础。 方法： 通过PCR扩增幽门螺杆菌CagA基因，克隆至腺病毒穿梭载体pAdTrack-CMV，该重组质粒与腺病毒DNA共转化E.coli BJ5183，通过细菌内同源重组获得重组腺病毒DNA，将其转染293细胞获得重组腺病毒。 结果和结论： 成功构建了幽门螺杆菌CagA的非复制型重组腺病毒。

Construction of recombinant adenoviruses expressing the helicobacter pylori CagA antigen

AIM: To construct a replication - defective reeombinant adenovirus, which expresses the CagA gene. METHODS AND RESULTS: The CagA gene was amplified by PCR. This heterogeneous gene was cloned into shuttle vector pAd-Track - CMV. The reeombinant adenovirus DNA was obtained by the homologous recombination between the shuttle vector and adenovirus DNA in E. coli 5183. After linearization, the reeombinant adenovirus DNA was transfected into 293 cells and the reeombinant adenovirus was obtained. Through this technique, the replication - defective reeombinant adenovirus AdEasyCagA was constructed. CONCLUSIONS: The replication- defective reeombinant adenoviruses AdEasyCagA was constructed successfully. The work will make a good foundation for studying the effects of the replication - defective reeombinant adenovirus on Th1/Th2 balance in asthma and be useful for finding a new pathway to prevent and cure asthma.