关键词: 尘螨抗原;基因;克隆;表达

    摘 要:目的 构建并克隆屋尘螨抗原Der p2基因，在大肠杆菌中初步表达. 方法 根据已发表的Der p2基因，人工设计合成一系列基因片段，经单链扩增后连接成完整的Der p2基因;将该基因克隆入表达载体pProEX HTb中，测序并进行初步表达. 结果 利用所设计的结构制备了完整的Der p2基因，以大肠杆菌为宿主在IPTG的诱导下获得初步表达，该外源蛋白Mr 约为17×103 ，占总蛋白量的20%. 结论 所构建的Der p2基因能在大肠杆菌中表达.

    Cloning and expression of gene of house dust mite antigen Der p2

    SHI Jie-Ran LI Yuan QI Hao-Wen LI Bie-Hu BAI Yin-Lan FAN Xiong-Lin XUE Ying

    1 Department of Respiratory Disease，Xijing Hospi-tal，

    2 Department of Microbiology，Faculty of Pre-clinical Medicine，Fourth Military Medical Universi-ty，Xi'an710033，China

    Keywords:dust mite antigen;gene;cloning;expression

    Abstract:AIM To construct and clone the gene of the house dust mite antigen Der p2，which is expressed in E.coli.METHODS The gene fragments were designed and synthesized by the published house dust mite antigen Der p2.Using single chain amplification by PCR and T4ligase，we prepared the whole Der p2gene，which was cloned to express vector pProEX HTb for sequence analysis and expression.RESULTS The whole Der p2gene was constructed with designed structure and expressed by IPTG inducement in E.coli.The Der p2protein was detected to be about Mr 17×103 by SDS-PAGE.The yield account for up to20%of the total amount of protein.CONCLUSION The constructed Der p2gene can be expressed in E.coli. ......