放射诱导启动子的合成与鉴定及质粒pcDNA3.1(+)/E-CDglyTK 的构建

【目的】合成放射诱导启动子序列并构建其调控双自杀基因的真核表达质粒pcDNA3.1 (+)/ECDglyTK 。【方法】利用人工寡核苷酸片段合成放射诱导启动子, 以绿色荧光蛋白(GFP) 作为报告基因转染 Tca8113 细胞, 经流式细胞仪鉴定其辐射诱导特性。将pCEA-CDglyTK 中双自杀基因CDglyTK 亚克隆到 pcDNA3.1(+), 然后把合成启动子插入到CDglyTK 上游构建质粒pcDNA3.1(+)/E-CDglyTK 并酶切鉴定, 阳离子 脂质体介导其转染舌鳞癌Tca8113 细胞, RT-PCR 观察CDglyTK 表达及诱导放疗的增效作用。【结果】合成启动 子序列...

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Published inZhongshan da xue xue bao. Zhongshan daxue xuebao yixue kexue ban = Journal of Sun Yat-sen University. Yi xue ke xue ban Vol. 26
Main Authors 余东升, 黄洪章, 潘朝斌, 王安训, 王建广
Format Journal Article
LanguageChinese
Published Editorial Office of Journal of Sun Yat-sen University 01.01.2005
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Abstract 【目的】合成放射诱导启动子序列并构建其调控双自杀基因的真核表达质粒pcDNA3.1 (+)/ECDglyTK 。【方法】利用人工寡核苷酸片段合成放射诱导启动子, 以绿色荧光蛋白(GFP) 作为报告基因转染 Tca8113 细胞, 经流式细胞仪鉴定其辐射诱导特性。将pCEA-CDglyTK 中双自杀基因CDglyTK 亚克隆到 pcDNA3.1(+), 然后把合成启动子插入到CDglyTK 上游构建质粒pcDNA3.1(+)/E-CDglyTK 并酶切鉴定, 阳离子 脂质体介导其转染舌鳞癌Tca8113 细胞, RT-PCR 观察CDglyTK 表达及诱导放疗的增效作用。【结果】合成启动 子序列分析与设计序列完全一致; 低剂量放射线照射可诱导这种人工启动子增强GFP 在Tca8113 细胞中的表 达; 重组质粒的酶切图谱与预期一致; 3 Gy 放疗显著增强CDglyTK 在Tca8113 细胞中的表达。【结论】成功合 成放射诱导启动子并构建由其调控双自杀基因的真核表达质粒pcDNA3.1(+)/E-CDglyTK, 为进一步研究肿瘤 放射- 基因治疗奠定了基础。
AbstractList 【目的】合成放射诱导启动子序列并构建其调控双自杀基因的真核表达质粒pcDNA3.1 (+)/ECDglyTK 。【方法】利用人工寡核苷酸片段合成放射诱导启动子, 以绿色荧光蛋白(GFP) 作为报告基因转染 Tca8113 细胞, 经流式细胞仪鉴定其辐射诱导特性。将pCEA-CDglyTK 中双自杀基因CDglyTK 亚克隆到 pcDNA3.1(+), 然后把合成启动子插入到CDglyTK 上游构建质粒pcDNA3.1(+)/E-CDglyTK 并酶切鉴定, 阳离子 脂质体介导其转染舌鳞癌Tca8113 细胞, RT-PCR 观察CDglyTK 表达及诱导放疗的增效作用。【结果】合成启动 子序列分析与设计序列完全一致; 低剂量放射线照射可诱导这种人工启动子增强GFP 在Tca8113 细胞中的表 达; 重组质粒的酶切图谱与预期一致; 3 Gy 放疗显著增强CDglyTK 在Tca8113 细胞中的表达。【结论】成功合 成放射诱导启动子并构建由其调控双自杀基因的真核表达质粒pcDNA3.1(+)/E-CDglyTK, 为进一步研究肿瘤 放射- 基因治疗奠定了基础。
Author 潘朝斌
王安训
黄洪章
王建广
余东升
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Snippet 【目的】合成放射诱导启动子序列并构建其调控双自杀基因的真核表达质粒pcDNA3.1 (+)/ECDglyTK 。【方法】利用人工寡核苷酸片段合成放射诱导启动子, 以绿色荧光蛋白(GFP) 作为报告基因转染 Tca8113 细胞, 经流式细胞仪鉴定其辐射诱导特性。将pCEA-CDglyTK 中双自杀基因CDglyTK...
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