| 颜冬菁1 盖文丽1 赵燕英2 黄欣媛1 陈正望1 陆婕1.小鼠Daintain/AIF-1 基因启动子的克隆及其荧光素酶报告基因
载体的构建[J].现代生物医学进展英文版,2011,11(6):1005-1008. |
| 小鼠Daintain/AIF-1 基因启动子的克隆及其荧光素酶报告基因
载体的构建 |
| Molecular Cloning of Daintain/AIF-1 Gene Promoter and Constructing ofLuciferase Report Gene Vector In Mice |
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| DOI： |
| 中文关键词: Daintain/AIF-1 基因启动子 荧光素酶报告基因载体 构建 |
| 英文关键词: Daintain/AIF-1 gene promoter Luciferase report gene vector Construction |
| 基金项目:湖北省自然科学基金（2009CDB076）；华中科技大学自主创新研究基金（2010MS009 ) |
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| 中文摘要: |
| 目的：对Daintain/AIF-1（大炎肽/ 同种异体移植炎症因子-1）基因启动子进行克隆并构建荧光素酶报告基因载体，为进一步
研究Daintain/AIF-1 的转录调控作用提供了质粒资源。方法：提取单核巨噬细胞系RAW264.7 基因组DNA，以其为模板采用PCR
方法克隆出Daintain/AIF-1 基因5' 端UTR 区1.6 kb DNA 序列，将该序列同源重组到pGL3-Basic 载体上，转化感受态DH5α 并
酶切鉴定和测序。结果：PCR 产物片段与预期结果一致，Daintain/AIF-1 基因5' 端UTR 区1.6 kb DNA 序列连接到pGL3-Basic 载
体上，构建成pGL3-Basic-Daintain/AIF-1 (pGL3-Basic-DT) 载体，酶切结果与理论预测值一致，经测序证实无碱基突变。结论：
Daintain/AIF-1 基因报告基因载体的构建为进一步研究Daintain/AIF-1 转录调控作用提供了载体资源。 |
| 英文摘要: |
| Objective: To construct luciferase report gene vector for mouse Daintain/AIF-1 gene promoter in order to provide
foundation for further study on the transcription regulation of Daintain/AIF-1. Methods: The mouse genome was extracted from the
macrophage cells RAW264.7, and from which 1.6 kb DNA sequence of Daintain/AIF-1 gene 5' end UTR was amplified by PCR. The
PCR product was directly inserted into pGL3-Basic vector by homologous recombination and the vector was transferred into DH5α. The
positive clone was identified by digestion with NcoⅠ and sequenced. Results: A 1.6 kb DNA sequence of Daintain/AIF-1 gene 5' end
UTRwas successfully cloned and the luciferase report gene vector ofmouseDaintain/AIF-1 gene promoterwaswell constructed. Conclusion:
pGL3-Basic-Daintain/AIF-1 (pGL3-Basic-DT) vector was constructed, which lays a foundation for further research on Daintain/AIF-1
gene transcription regulation. |
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