Mamu-B* 007:03及其剪切异构体Mamu-B* 007:03-sv1基因的表达与细胞内定位

Expression and cellular localization of and its splice variant Mamu-B* 007:03 Mamu-B* 007:03-sv1

目的 Mamu-B*007:03-sv1是中国恒河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α3结构域缺失。本实验初步获得其在细胞内的表达和定位信息,并与全长的Mamu-B*007:03基因的表达情况进行比较。方法分别构建Mamu-B*007:03-sv1-myc-pEGFPN3和Mamu-B*007:03-myc-pEGFPN3的真核表达载体,并将这两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两种基因在细胞内的表达情况,并利用激光共聚焦显微镜技术分别分析这两种基因在细胞内的表达定位。结果 Mamu-B*007:03-sv1和Mamu-B*007:03基因均能在293T细胞内正常表达。Mamu-B*007:03-sv1发生了糖基化的修饰,Mamu-B*007:03基因在细胞膜上表达,Mamu-B*007:03-sv1基因在细胞内表达。结论 Mamu-B*007:03-sv1基因的α3结构域缺失,其细胞内的表达和定位与全长Mamu-B*007:03基因有明显差异,其功能有待于进一步探索。

Objective Mamu-B ＊ O07：03-svl is a splice variant of major histocompatibility complex class I in Chinese rhesus macaque devoid of α3 domain. The aim of this study was to get the basic information on expression and cellular localization of Mamu-B ＊ 007：03-svl and to compare with the full-length Mamu-B ＊ 007 ： 03 gene. Methods Eukaryotic expression vectors of Mamu-B ＊ 007： 03-svl-myc-pEGFPN3 and Mamu-B ＊ 007： 03-myc-pEGFPN3 were constructed and transfected into human renal epithelial 293T cells. The expression of the two genes in cells was detected by Western blotting, and the cellular localization of the two genes was analyzed by immunofluorescence and laser scanning confocal microscopy. Results Both Mamu-B ＊ 007：03-svl and Mamu-B ＊ 007：03 were expressed in 293T cells and both were glycosylated. While Mamu-B ＊ 007：03 gene was expressed on the cell membrane, Mamu-B ＊ 007：03-svl gene was expressed intracellularly and on cell membrane. Conclusions The expression and cellular localization of Mamu-B ＊ 007 ： 03-svl deviod of α3 is significantly different from that of the full length Mamu-B ＊ 007：03 gene. The functions of Mamu-B ＊ 007：03-svl remain to be further studied.