| 张翠珍1 蒋学俊1△ 钱航2 张鹏2 杨汉东2△.含G0S2 基因启动子的荧光素酶报告基因载体的构建*[J].,2014,14(10):1830-1833 |
| 含G0S2 基因启动子的荧光素酶报告基因载体的构建* |
| Construction and Significance of G0S2 Promoter-DirectedLuciferase Reporter Gene Plasmid* |
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| DOI： |
| 中文关键词: G0S2 基因 启动子 荧光素酶 报告基因质粒 |
| 英文关键词: G0S2 Promoter Luciferase Reporter gene vector |
| 基金项目:湖北省教育厅科研项目（T201212 和Q20122401） |
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| 中文摘要: |
| 摘要目的：克隆人G0S2 基因启动子并构建荧光素酶报告基因载体，为进一步研究G0S2 基因转录调控提供质粒。方法：利用
PCR 技术从人胚肾293A 细胞基因组DNA 中克隆获得G0S2 基因启动子的DNA 片段，将其克隆至pGL3-basic 表达载体中，并
转化人大肠杆菌DH5琢，经限制性内切酶酶切、PCR 及测序鉴定得到确认；将重组载体质粒与半乳糖苷酶表达质粒
psV-茁-Galactosidase 共转染至大鼠血管平滑肌细胞（VSMC），检测细胞中荧光素酶的活性。结果：pGL3-G0S2-Promoter 重组质粒
插入片段和相邻序列正确，克隆的G0S2 基因片段有启动子活性（P<0.05）。结论：成功构建了pGL3-G0S2-Promoter 报告基因质
粒，为进一步研究G0S2 基因的表达奠定了基础。 |
| 英文摘要: |
| ABSTRACT Objective：To clone the luciferase reporter gene and construct promoter region from human G0S2 gene. Methods：A
region containing human G0S2 gene promoter was obtained by PCR amplification from 293A cells, and the segment was cloned into the
pGL3-Basic vector and transformed into , which was verified by endonuclease, PCR amplification and direct sequencing. The final
construct (pGL3-G0S2-Promoter) was transferred into VSMC (vascular smooth muscle cells), and the luciferase activity was measured.
Results：The insert and surrounding regions in pGL3-G0S2-Promoter plasmid were confirmed. There was significant promoter activity for
pGL3-G0S2-Promoter vector (P<0.05).Conclusion：Luciferase reporter plasmid pGL3-G0S2-Promoter containing G0S2 promoter can
be built successfully. It will be used to study the transcriptional regulation of G0S2 gene. |
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