棕头蜘蛛猴Atfu-B基因α2结构域缺失剪切异构体的表达及其细胞内定位

Expression and intracellular localization of splice variants of Ateles fusciceps gene Atfu-B with α2 domain deletion

目的 Atfu-B* sv1是棕头蜘蛛猴主要组织相容性复合体(major histocompatibility complex,MHC)I类分子的一种剪切异构体,其α2结构域缺失。本实验旨在初步获得其在细胞内的表达和定位信息,并与全长的Atfu-B基因的表达情况进行比较,观察两者之间的差异。方法分别构建Atfu-B*sv1以及全长Atfu-B基因AtfuB*02:03和Atfu-B*03:01的真核表达载体,并将这三种重组质粒分别转染293T细胞。利用Western blot免疫印迹实验检测这三种基因在细胞内的表达情况,并利用免疫荧光和激光共聚焦显微镜技术分别分析这三种基因在细胞内的表达定位。结果 Atfu-B*sv1、Atfu-B*02:03和Atfu-B*03:01均能在293T细胞内正常表达,并且都发生了糖基化的修饰,三种基因都在细胞膜上表达。结论虽然Atfu-B*sv1的α2结构域缺失,但其细胞内的表达和定位与全长全长Atfu-B基因没有明显差异,其功能有待于进一步探索。

Objective Affu-B ＊ svl with α2 domain deletion is a splice variant of major histocompatibility complex class I B in Ateles fuseieeps. The aim of this study was to obtain the basic information of expression and intracellular localization of Atfu-B＊svl, and to compared them with that of the full-length Affu-B gene. Methods Eukaryotic expression vectors of Affu-B＊svl, Affu B＊02：03 and Affu-B＊03：01 were constructed and transfected into 293T cells, respectively. Expression of the three genes in the cells was detected by Western blotting, and the intracellular localization of the three genes was analyzed by immunofluorescence and laser confocal microscopy. Results Atfu-B,svl, Atfu-B,02：03 and Atfu- B＊03：01 were all expressed on the plasma membrane of 293T cells and were glycosylated. Conclusions The expression and intracellular localization of Atfu-B＊svl is not significantly different from those of full length Atfu-B genes. Further studies on the functions of Atfu-B＊svl are needed to be done.