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构建由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腺病毒 被引量：3

Construction of replication-defective recombinant adenovirus with FCY1 driven by multimer of hypoxia responsive elements-modified hTERT core promoter
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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 (h TERT)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 h TERT启动子上游并经测序证实 .将修饰后的 h TERT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 HEK2 93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 h TERT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 / lox P的腺病毒载体构建系统操作简便。 AIM To construct replication defective recombinant adenovirus with FCY1 driven by multimer of hypoxia responsive elements modified hTERT core promoter. METHODS Multimer of hypoxia responsive elements were designed and cloned upstream to the hTERT core promoter, then confirmed by sequencing. The modified hTERT promoter and FCY1 were inserted into shuttle plasmids and cotransfected HEK 293 cells with rescue plasmid pBHGlox (delta) E1, 3Cre. RESULTS Recombinant adenoviruses with FCY1 driven by three or six copies of hypoxia responsive elements modified hTERT core promoter were constructed and amplified. The titers of the resulting adenoviruses were 8.4×10 10 pfu·L -1 and 6.9×10 10 pfu·L -1 , respectively, as determined by end point dilution assay. CONCLUSION The adenoviral vector construction kit based on Cre recombinase/loxP system can be easily and efficiently used in construction of replication defective adenovirus.
作者 李立文 王臻 苏明权 马越云 于文彬 杨旻 李哲
机构地区 第四军医大学西京医院全军骨科研究所 第四军医大学西京医院临床分子生物学研究室
出处 《第四军医大学学报》 北大核心 2002年第12期1061-1064,共4页 Journal of the Fourth Military Medical University
基金 国家自然科学基金资助 (3 0 170 95 2 )
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 anoxia response elements transcriptional targeting telomerase adenoviridae genetic vectors
分类号 R730 [医药卫生—肿瘤]
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参考文献20

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同被引文献19

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第四军医大学学报
2002年 第12期

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