尤红娟1 郝林2 梁清2 贺厚光2 刘雯1 刘转转1 汤仁仙1 韩从辉2△.徐州医学院病原生物学与免疫学教研室[J].现代生物医学进展英文版,2012,12(20):3835-3837. |
徐州医学院病原生物学与免疫学教研室 |
Construction and Evaluation of the Luciferase Reporter Vectorwith the PSA Promoter |
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DOI: |
中文关键词: PSA 启动子 荧光素酶报告载体 克隆 |
英文关键词: PSA promoter Luciferase reporter vector Clone |
基金项目:江苏省分子和功能影像重点实验室开放基金(PYZX2011001);徐州市医学科研项目(XWJ2011027) |
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中文摘要: |
目的:建立高活性的PSA 启动子荧光素酶报告载体。方法:提取前列腺癌细胞PC-3 总DNA,PCR 扩增出PSA 启动子片段,
建立PSA 启动子荧光素酶重组载体PGL3-psap,通过脂质体介导将重组载体转染到前列腺癌细胞PC-3 中,使用荧光素酶检测系
统检测其活性。结果:DNA 测序结果表明成功构建荧光素酶重组载体PGL3-psap,荧光素酶检测显示重组载体在前列腺癌细胞
PC-3 中有较强活性。结论:本研究成功构建了具有高度活性的PSA 启动子荧光素酶报告载体,为进一步研究前列腺癌的诊断和
治疗提供了实验基础。 |
英文摘要: |
Objective: To clone PSA gene promoter and construct PGL3-basic luciferase reporter vector with PSA promoter, and
analysis its activity in PC-3. Methods: To design and amplify PSA gene promoter, then clone the gene into PGL3-basic, and obtain
luciferase reporter vectors PGL3-psap. To compare the luciferase activity of cell lines transiently transfected by PGL3-psap, PGL3-Basic
and pGL3-control. Results: The PSA gene promoter was obtained by PCR. The DNA sequencing analysis showed that the luciferase
reporter vector with PSA gene promoter had been constructed successfully. The cell lines were transiently transfected by PGL3-psap,
PGL3-Basic and pGL3-control. The luciferase activity of the group increased after transiently transfected by PGL3-psap. Conclusion: The
luciferase reporter vectors with PSA promoters had been successfully constructed,so we supplied experimental basis for diagnosis and
treatment of prostatic cancer. |
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