携带细胞周期调控基因p14~(ARF)的重组腺病毒的构建
Construction of Recombinant Adenovirus Vector Carrying Cell Cycle Controlling Gene-p14ARF
作者：鲜均明;周光耀;梁传余;刘世喜
Author：XIAN Jun-ming1, ZHOU Guang-yao2, LIANG Chuan-yu2, LIU Shi-xi2
收稿日期：2006-06-25 年卷（期）页码：2006,24(03):261-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：细胞信号转导,重组腺病毒,基因治疗,喉癌,
Key words：cell signal transduction,recombinant adenovirus, gene therapy,laryngeal squamous cell carcinoma,
基金项目：
国家自然科学基金资助项目（0040205401078）；四川省青年科技基金资助项目（04ZQ026-024）
中文摘要
目的 构建携带参与细胞周期调控的p14ARF基因的重组腺病毒载体。方法 将野生型p14ARF全长cDNA基因片段插入腺病毒载体质粒pAdTrack-CMV，与骨架质粒在大肠杆菌BJ5183胞内进行同源重组，经293细胞包装、扩增后得到携带p14ARF的重组腺病毒AdEasy-GFP-p14ARF。结果 成功构建了AdEasy-GFP-p14ARF的重组腺病毒载体系统，经测定腺病毒载体滴度达2.3×109 efu/mL。结论 构建的重组腺病毒AdEasy-GFP-p14ARF可望有效地将p14ARF基因导入喉癌细胞株/组织内，为进一步研究喉癌细胞信号转导干预治疗机制提供实验基础。
英文摘要
Objective The recombinant adenovirus vector carrying p14ARF gene was constructed for using in the interference therapy in signal transduction of laryngeal squamous cell carcinoma. Methods The total cDNA fragment of p14ARF was cloned into the shuttle plasmid pAdTrack-CMV, with the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the E.Coli BJ5183 and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged and amplified in the 293 cells. Then the viral titer was checked by GFP. Results The recombinant adenovirus vector carrying p14ARF was constructed successfully. The viral titer was 2.3×109. Conclusion The recombinant adenovirus vector could introduce p14ARF gene into the laryngeal squamous cell carcinoma line or tumor tissue effectively, which would provide experimental basis for the mechanisms and further study of the interference therapy in signal transduction of laryngeal squamous cell carcinoma.
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