电压门控快钾通道蛋白Kv4.3的抗体制备及检测  

The Antiserum of the Multiple Antigenic Peptide of the Kv4.3:Preparation and Applicat ion

作　　者：邓小云[1] 刘宇[1] 蔡芳[1] 潘乾[1] 龙志高[1] 夏昆[1] 夏家辉[1] 张灼华[1] 

机构地区：[1]中南大学医学遗传学国家重点实验室,中国湖南长沙410078

出　　处：《生命科学研究》2005年第1期6-10,共5页Life Science Research

基　　金：国家自然科学基金资助项目(30170454;30123006;30300101)

摘　　要：Kv4(voltagegatedK+channel4)是在哺乳动物心脏和脑组织中广泛表达的一类钾离子通道蛋白.它主要介导心肌细胞和神经细胞中的A-型(快速失活型)K+电流,是构成中枢神经系统和心肌细胞中的快速失活外向型电流的基础.Kv4.3是电压门控钾离子通道3种α亚基之一.通道中含有许多具有调节作用的辅助亚基,它们通过与Kv4.3互作实现对通道的调节.本研究根据人类氨基酸序列以MAPs法制备抗Kv4.3抗体.MAP由4条相同的17肽段连接成锥形结构分子.以MAP或MAP与佐剂免疫新西兰白兔,抽提了免疫前血清和免疫后血清,并对抗体的滴度进行评价.MAPs在白兔体内诱导出了效价比为1:1000的抗Kv4.3特异性抗体.Voltage-dependent potassium chan nel proteins are highly expressed in the heart and the brain of mammal,which mediates the A-type cu rrent in the neuronal cell and cardia c myocytes.Kv4.3is one of the threeαsubunits of voltage-dependent pota ssium channel proteins.Many Kv chan nels contain auxiliary subunits that can regulate the biophysical,b iochemical,and cell biological characteristics of the resultant channel complexes.In this study the anti-Kv 4.3antibody was raised by multiple a ntigenic peptide solution.The MAP was constructed of a four-spiked rin g.Following vaccinations with the MAP or MAP plus adjuvant,sera were taken and antibody titres were assessed.Further analysis also demonstr ated that these peptides were able to generate antibody responses that re cognize rattus Kv4.3sequences.

关 键 词：Kv4.3 MAPS WESTERN BLOTTING 免疫沉淀 

分 类 号：Q31[生物学—遗传学]