| 江冠民，匡艳华，邱瑜，，谢婉莹，张秋桂，欧阳新平4.IDO启动序列GAS和ISRE荧光素酶报告基因载体构建及其活性检测.[J].中南医学科学杂志.,2014,42(2):120-124. |
| IDO启动序列GAS和ISRE荧光素酶报告基因载体构建及其活性检测 |
| Construction and Activity Assay of Luciferase Reporter VectorContaining Promoter Sequence GAS and ISRE of IDO |
| 投稿时间：2013-11-10 |
| DOI： |
| 中文关键词: 吲哚胺2，3-双加氧酶 启动子序列 荧光素酶报告基因 γ-干扰素 肿瘤免疫耐受 |
| 英文关键词:indoleamine 2,3-dioxygenase promoter sequence luciferase reporter gene IFN-γ tumor immune tolerance |
| 基金项目:湖南省自然科学基金资助项目（13JJ4078). |
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| 中文摘要: |
| 目的构建pGL3-Enhancer-ISRE4和pGL3-Enhancer-GAS两个荧光素酶报告基因载体，并对其进行生物活性鉴定。方法通过人工合成调控吲哚胺2，3-双加氧酶（IDO）表达的启动序列ISRE4（4个串联的ISRE序列）和GAS7（7个串联的GAS序列），与pGL3-Enhancer连接成重组体pGL3-Enhancer-ISRE4和pGL3-Enhancer-GAS7，通过转化扩增，筛选出阳性克隆，并通过酶切、测序及生物学活性检测鉴定构建好的荧光素酶报告基因载体。 |
| 英文摘要: |
| ObjectiveTo construct two luciferase reporter gene vector pGL3-Enhancer-ISRE4 and pGL3-Enhancer-GAS7,and to identify their biological activity of them.MethodsThe promoter sequences ISRE4 (4 series ISRE sequence) and GAS7 (7 series GAS sequence) were synthesized which regulate the expression of IDO,then they were cloned into empty vector pGL3-Enhancer respectively to construct two luciferase reporter vectors pGL3-Enhancer-ISRE4 and pGL3-Enhancer-GAS7,which were amplified by transformation and identified by restriction enzyme digestion,sequencing,and the biological activity detecting.ResultsTwo luciferase reporter vectors pGL3-Enhancer-ISRE4 and pGL3-Enhancer-GAS7 were successfully constructed,the expression of luciferase could be induced by IFN-γ.ConclusionThe two luciferase reporter gene vectors of pGL3-Enhancer-ISRE4 and pGL3-Enhancer-GAS provide a very important tool for studying the regulation mechanisms of IDO protein expression and supplying a fast,high-throughput screening for anti-tumor immune tolerance drug which targeting the IDO. |
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