CAJ | 학술논문

目的根据前期筛选到的与ataxin-3蛋白羧基端相互作用的A3IP部分序列,进行A3IP基因的克隆,并探讨其细胞及组织定位情况.方法应用生物信息学及Northern印迹方法克隆A3IP基因并检测其转录本在人体组织和人脑不同部位的表达情况；应用Western印迹及免疫荧光方法检测A3IP蛋白在细胞中的表达及定位情况；应用免疫组织化学染色方法检测A3IP蛋白在人体各组织和人脑不同部位的表达及定位情况.结果对A3IP基因序列全长阅读框进行了cDNA克隆及序列拼接；检测了A3IP的3个转录本(1 kb、1.35 kb、6 kb)在人体不同组织和人脑不同部位的表达谱；获得了A3IP-pEGFP在COS-7细胞和人大脑皮质、小脑、肌肉、周围神经、肝脏、肾脏的表达及定位情况.结论所克隆的A3IP基因编码ataxin-3的一种相互作用蛋白A3IP,其3个剪切本在人体多种组织和人脑不同部位广泛表达,是一种细胞胞浆蛋白.
목적 근거전기사선도적여ataxin-3단백최기단상호작용적A3IP부분서렬,진행A3IP기인적극륭,병탐토기세포급조직정위정황.방법 응용생물신식학급Northern인적방법극륭A3IP기인병검측기전록본재인체조직화인뇌불동부위적표체정황；응용Western인적급면역형광방법검측A3IP단백재세포중적표체급정위정황；응용면역조직화학염색방법검측A3IP단백재인체각조직화인뇌불동부위적표체급정위정황.결과 대A3IP기인서렬전장열독광진행료cDNA극륭급서렬병접；검측료A3IP적3개전록본(1 kb、1.35 kb、6 kb)재인체불동조직화인뇌불동부위적표체보；획득료A3IP-pEGFP재COS-7세포화인대뇌피질、소뇌、기육、주위신경、간장、신장적표체급정위정황.결론 소극륭적A3IP기인편마ataxin-3적일충상호작용단백A3IP,기3개전절본재인체다충조직화인뇌불동부위엄범표체,시일충세포포장단백.
Objective To clone an A3IP gene and investigate its cellular and histological localization based on previous research which has identified part of A3IP sequence interacting with carboxyl-terminal of ataxin-3.Methods Bioinformatic and Northern blotting were applied to clone the A3IP gene and detect the expression of its transcripts in various human tissues and brain regions.Western blotting and immunofluorescence staining were applied to detect expression of A3IP protein in cultured cells.Immunohistochemistry staining was applied to study the expression of A3IP protein in various human tissues and brain regions.Results cDNA cloning of A3IP gene's reading frame and its sequence assembly were completed.Three transcripts (1 kb,1.35 kb and 6 kb,respectively) of A3IP were found to express in various human tissues and brain regions.A3IP-pEGFP expresses in cytoplasm of cultured COS-7 cells and various human tissues and brain regions including cerebral cortex,cerebellum,muscle,peripheral nerve,liver and kidney.Conclusion The cloned A3IP gene encodes A3IP,a novel ataxin-3-interacting protein.Three transcripts of A3IP are expressed in various human tissues and brain regions.A3IP is a cytosolic protein.