Mamu-B*007:03及其剪切异构体Mamu-B*007:03-sv1基因的表达及细胞内定位研究
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Expression and cellular localization of Mamu-B*007:03-sv1 and Mamu-B*007:03
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    摘要:

    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。

    Abstract:

    Objective Mamu-B*007:03-sv1 is a splice variant of major histocompatibility complex class I in Chinese rhesus macaque devoid of α3 domain. In this experiment the basic information about expression and cellular location of Mamu-B*007:03-sv1 was got and compared with the full-length Mamu-B*007:03 genes. Methods Eukaryotic expression vectors of Mamu-B*007:03-sv1-myc-pEGFPN3 and Mamu-B*007:03-myc-pEGFPN3 were constructed and transfected to 293T cells. The expressions of the two genes in cells were detected by Western blotting methods, and the cellular locations of the two genes were analyzed by immunofluorescence and laser confocal microscopy. Results Mamu-B*007:03-sv1 and Mamu-B*007:03 were all expressed in the 293T cell and were all glycosylation. While Mamu-B*007:03 gene was expressed on the cell membrane, and Mamu-B*007:03-sv1 gene was expressed in both intra-cellular and cell membrane. Conclusion The expression and cellular localization of Mamu-B*007:03-sv1 deviod of α3 was significantly different from the full length Mamu-B*007:03 gene. Further researches about functions of Mamu-B*007:03-sv1 need to be done.

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严翔. Mamu-B*007:03及其剪切异构体Mamu-B*007:03-sv1基因的表达及细胞内定位研究[J].中国比较医学杂志,2013,23(12).

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  • 收稿日期:2013-10-12
  • 最后修改日期:2013-10-18
  • 录用日期:2013-11-26
  • 在线发布日期: 2014-02-11
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