关键词: 低氧;反应元件;转录靶向;端粒，末端转移酶;腺病毒科;遗传载体

    摘 要:目的 构建由缺氧反应元件串联重复体修饰的人端粒酶逆转录酶(hTERT)核心启动子引导酵母胞嘧啶脱氨酶FCY1基因表达的复制缺陷型重组腺病毒. 方法 人工设计缺氧反应元件串联重复序列，克隆后插入hTERT启动子上游并经测序证实.将修饰后的hTERT启动子和酵母胞嘧啶脱氨酶基因FCY1插入穿梭质粒，与辅助质粒共转染HEK293细胞，重组产生复制缺陷型腺病毒. 结果 获得了由3倍和6倍缺氧反应元件修饰的hTERT核心启动子引导FCY1基因表达的复制缺陷型重组腺病毒. 结论 基于Cre重组酶/loxP的腺病毒载体构建系统操作简便、重组效率高.

    Construction of replication┐defective recombinant adenovirus with FCY1driven by multimer of hypox┐ia responsive elements┐modified hTERT core pro-moter

    LI Li-Wen WANG Zhen SU Ming-Quan MA Yue-Yun YU Wen-Bin YANG Min LI Zhe

    1 Institute of Orthopaedics of Chinese PLA，

    2 Laboratory of Clinical Molecular Biology，Xijing Hospital，Fourth Military Medical University，Xi'an710033，China

    Keywords:anoxia;response elements;transcriptional target-ing;telomerase;adenoviridae;genetic vectors

    Abstract:AIM To construct replication-defective recombi-nant adenovirus with FCY1driven by multimer of hypoxia re-sponsive elements-modified hTERT core promoter.METH┐ODS Multimer of hypoxia responsive elements were de-signed and cloned upstream to the hTERT core promoter ......