大鼠肝脏间隙连接蛋白cDNA的分子克隆
Molecular cloning of cDNA for rat liver gap junction protein.
作者信息
Paul D L
出版信息
J Cell Biol. 1986 Jul;103(1):123-34. doi: 10.1083/jcb.103.1.123.
Abstract
An affinity-purified antibody directed against the 27-kD protein associated with isolated rat liver gap junctions was produced. Light and electron microscopic immunocytochemistry showed that this antigen was localized specifically to the cytoplasmic surfaces of gap junctions. The antibody was used to select cDNA from a rat liver library in the expression vector lambda gt11. The largest cDNA selected contained 1,494 bp and coded for a protein with a calculated molecular mass of 32,007 daltons. Northern blot analysis indicated that brain, kidney, and stomach express an mRNA with similar size and homology to that expressed in liver, but that heart and lens express differently sized, less homologous mRNA.
摘要
制备了一种针对与分离的大鼠肝脏缝隙连接相关的27-kD蛋白的亲和纯化抗体。光镜和电镜免疫细胞化学显示该抗原特异性定位于缝隙连接的细胞质表面。该抗体用于从表达载体λgt11中的大鼠肝脏文库中筛选cDNA。筛选出的最大cDNA包含1494 bp，编码一种计算分子量为32007道尔顿的蛋白质。Northern印迹分析表明，脑、肾和胃表达的mRNA大小和同源性与肝脏中表达的相似，但心脏和晶状体表达的mRNA大小不同且同源性较低。
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