人CYP19基因启动子荧光素酶报告基因载体的构建 |
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引用本文: | 邵喜英,陈占红,黄健,雷蕾,冯建国,苏丹,王晓稼. 人CYP19基因启动子荧光素酶报告基因载体的构建[J]. 肿瘤学杂志, 2010, 16(11): 873-876 |
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作者姓名: | 邵喜英 陈占红 黄健 雷蕾 冯建国 苏丹 王晓稼 |
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作者单位: | [1]浙江省肿瘤医院,浙江杭州310022 [2]浙江省肿瘤研究所,浙江杭州310022 |
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基金项目: | 浙江省自然科学基金,浙江省医药卫生优秀青年科技人才专项基金,浙江省医药卫生科学研究基金 |
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摘 要: | [目的]克隆人CYP19基因启动子PⅠ.3(-345~-234bp)、PⅡ(-517~-278bp)和PⅠ.7(-299~+81bp)片段,构建启动子荧光素酶报告基因载体。[方法]采用生物合成PⅠ.3(-345~-234bp)、PⅡ(-517~-278bp)和PⅠ.7(-299~+81bp)区域基因片段,通过HindⅢ和KpnⅠ酶切连接到pGL3-Basic载体上。[结果]测序验证pGL3-Basic-PⅠ.3,pGL3-Basic-PⅡ和pGL3-Basic-PⅠ.7构建成功。[结论]pGL3-Basic-PⅠ.3,pGL3-Basic-PⅡ和pGL3-Basic-PⅠ.7荧光素酶报告基因载体的成功构建,为进一步研究CYP19基因组织特异性表达奠定基础。
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关 键 词: | CYP19基因 启动子 荧光素酶 |
Construction of Luciferase Report Gene Vector Containing Human CYP19 Promotor |
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Affiliation: | SHAO Xi-ying,CHEN Zhan-hong,HUANG Jian,et al.(Zhejiang Cancer Hospital,Hangzhou 310022,China) |
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Abstract: | [Purpose] To construct luciferase report gene vector containing human CYP19 promotor PⅠ.3(-345--234bp),PⅡ(-517--278bp)and PⅠ.7(-299-+81bp).[Methods] PⅠ.3(-345-234bp),PⅡ(-517--278bp)and PⅠ.7(-299-+81bp) were amplified by biosynthesis,and inserted into the luciferase report gene pGL3-Basic vector with restriction endonuclease HindⅢ and Kpn Ⅰ.[Results] pGL3-Basic-PⅠ.3,pGL3-Basic-PⅡ and pGL3-Basic-PⅠ.7 were confirmed by DNA sequencing.[Conclusion] The human CYP19 promotor luciferase reporter gene vectors are constructed successfully,which establishes the base for the further research of CYP19 tissue-specific expression mechanism. |
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Keywords: | CYP19 gene promotor luciferase |
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