人Tudor-SN UTR片段荧光素酶报告基因表达质粒的构建及活性检测

发布时间：2018-05-04 17:32

  本文选题：人Tudor-SN蛋白 + 　； 参考：《山东医药》2017年04期

【摘要】：目的为深入研究Tudor-SN蛋白本身在翻译水平的调控机制奠定基础。方法以HeLa细胞全基因组DNA为模板,PCR扩增出目的基因,利用双酶切的方法将目的片段连接到pGL3-Control载体上。再将构建成功的pGL3-5'UTR、pGL3-3'UTR重组质粒分别与内参海肾荧光素酶质粒瞬时共转染HeLa细胞,培养48h检测荧光素酶的活性。结果双酶切和基因测序法鉴定重组质粒构建成功,转染重组质粒后可检测到荧光素酶的活性;以pGL3-5'UTR质粒荧光素酶活性最高。结论成功构建了人Tudor-SN基因5'UTR和3'UTR序列的重组质粒,为研究Tudor-SN基因UTR区对翻译调控的影响奠定了基础。
[Abstract]:Objective to lay a foundation for further study on the regulation mechanism of Tudor-SN protein in translation level. Methods the target gene was amplified from the whole genomic DNA of HeLa cells and ligated to the pGL3-Control vector by double enzyme digestion. Then the constructed recombinant plasmid pGL3-5 UTR-pGL3-3 UTR was cotransfected into HeLa cells at the same time, and the luciferase activity was detected after 48 h culture. Results the recombinant plasmid was successfully constructed by double enzyme digestion and gene sequencing. The luciferase activity of the recombinant plasmid was detected after transfection, and the luciferase activity of the pGL3-5'UTR plasmid was the highest. Conclusion Recombinant plasmids of 5'UTR and 3'UTR of human Tudor-SN gene were successfully constructed, which laid a foundation for studying the effect of UTR region of Tudor-SN gene on translation regulation.
【作者单位】： 天津医科大学;
【基金】：国家杰出青年基金资助项目(31125012) 教育部“创新团队发展计划”(IRT13085) 国家自然科学基金资助项目(31170830/31370749/31571380) 天津市应用基础与前沿技术研究计划(青年基金项目)(15JCQNJC09900) 天津医科大学青年基金项目(2015KYZQ03)
【分类号】：Q78
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本文编号：1843932