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A split luciferase complementation assay for studying in vivo protein-protein interactions in filamentous ascomycetes

机译：分裂荧光素酶互补测定法，用于研究丝状子囊藻体内的蛋白质间相互作用

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摘要

The present invention sasangseong ascus for this vector and in vivo protein interaction analysis in fungi Using relates to a split luciferase complementation analysis method, and more particularly, to analyze whether the protein interaction sasangseong ascus in vivo from within the fungus, the protein expression vector the gene of interest After insertion, the split will luciferase assay and relates to a new vector that is used for this analysis, whether the protein interactions in vivo through a la kinase analysis. vector for protein interaction analysis of sasangseong ascus bacteria of the present invention is a gene encoding the N- terminal or C- terminal of the firefly luciferase; Creative parin gene ( Crp) of the promoter; EcoRI and SacI restriction enzyme sites; Geneticin (geneticin) or hygromycin B (hygromycin B) resistance gene cassette; And and HindIII ; and a restriction enzyme site (see Fig. 1). Further, a method for analyzing the interaction between the protein in bacteria sasangseong ascus in accordance with a preferred embodiment of the present invention is expressed by the split luciferase analysis by inserting the gene encoding the protein analyte in claim 1. Vector By measuring the fluorescence intensity of luciferase, sasangseong ascus is a method of analyzing the interaction between the protein in bacteria, characterized in that. ;

机译：用于该载体的本发明的三生曲霉和真菌中的体内蛋白质相互作用分析Using涉及分裂荧光素酶互补分析方法，更具体地讲，从内部分析了三生曲霉体内的蛋白相互作用。真菌，蛋白质表达载体感兴趣的基因插入后，拆分将进行荧光素酶测定，并涉及一种用于该分析的新载体，该蛋白质是否通过la激酶分析在体内相互作用。本发明的三生种曲霉细菌的蛋白质相互作用分析的载体是编码萤火虫荧光素酶的N末端或C末端的基因。启动子的创造性Parin基因（ Crp）; EcoRI 和 SacI 限制酶位点;遗传霉素（geneticin）或潮霉素B（hygromycin B）抗性基因盒;还有和HindIII ;和一个限制酶位点（见图1）。此外，通过将编码权利要求1的蛋白质分析物的基因插入其中，通过分裂萤光素酶分析来表达根据本发明优选实施方案的用于分析萨桑城曲霉中的蛋白质之间的相互作用的方法。载体通过测量荧光强度萤光素酶是三色as的一种分析细菌中蛋白质之间相互作用的方法，其特征在于。 ; 展开▼

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公开/公告号KR101460246B1

专利类型

公开/公告日2014-11-20

原文格式PDF

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申请/专利号KR20120059925

发明设计人 이승훈;윤성환;김희경;조은지;성보름;조성미;
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申请日2012-06-04

分类号C12N15/80;C12N1/15;C12Q1/66;

国家 KR

入库时间 2022-08-21 15:01:21

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1. 研究体内蛋白质相互作用的方法 [P] . 中国专利： CN1160470C . 2004.08.04

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3. VECTOR FOR ANALYZING PROTEIN INTERACTION IN ASSURGENCY ASCOMYCETE AND SPLIT LUCIFERASE COMPLEMENTATION ANALYZING METHOD USING SAME [P] . 韩国专利： KR20130136252A . 2013-12-12

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4. In vivo screening of protein-protein interactions with protein-fragment complementation assays [P] . 美国专利： US7855167B2 . 2010-12-21

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