胡建国, 王凤超, 钟政荣, 陆佩华. 大鼠GAP-43基因的克隆及其在COS-7细胞的表达[J]. 蚌埠医科大学学报, 2007, 32(3): 253-255.
    引用本文: 胡建国, 王凤超, 钟政荣, 陆佩华. 大鼠GAP-43基因的克隆及其在COS-7细胞的表达[J]. 蚌埠医科大学学报, 2007, 32(3): 253-255.
    HU Jian-guo, WANG Feng-chao, ZHONG Zheng-rong, LU Pei-hua. Cloning of rat GAP-43 gene and its expression in COS-7 cells[J]. Journal of Bengbu Medical University, 2007, 32(3): 253-255.
    Citation: HU Jian-guo, WANG Feng-chao, ZHONG Zheng-rong, LU Pei-hua. Cloning of rat GAP-43 gene and its expression in COS-7 cells[J]. Journal of Bengbu Medical University, 2007, 32(3): 253-255.

    大鼠GAP-43基因的克隆及其在COS-7细胞的表达

    Cloning of rat GAP-43 gene and its expression in COS-7 cells

    • 摘要: 目的: 克隆大鼠GAP-43基因,构建其真核表达载体,并观察其在哺乳动物细胞COS-7中的表达。方法: 采用RT-PCR方法,以大鼠少突胶质前体细胞RNA为模板,扩增GAP-43基因,定向克隆到pEGFP-N3载体中。以LipofectamineTM 2000试剂转染pEGFP-N3-GAP-43表达载体至COS-7细胞中进行瞬时真核表达。以免疫荧光方法鉴定GAP-43的表达。结果: 从大鼠少突胶质前体细胞中克隆到序列正确的GAP-43全长编码序列。所构建的GAP-43质粒在COS-7细胞中获得高效表达。结论: 大鼠GAP-43基因的克隆、真核表达载体的构建及在COS-7中的表达获得成功,为进一步研究其功能,尤其是探讨其在少突胶质细胞发育中的作用奠定了基础。

       

      Abstract: Objective: To clone rat GAP-43 gene and express it in mammalian cells.Methods: RT-PCR was used to amplify rat GAP-43 gene from RNA of rat oligodendrocyte precursor cells(OPCs).The GAP-43 gene was inserted into eukaryotic expression vector pEGFP-N3.The recombinant expression vector was transiently transfected into COS-7 cells by LipofectamineTM 2000 reagent.The expression of GAP-43 in COS-7 cells was detected by immunocytochemistry.Results: The sequence of the cloned GAP-43 was confirmed to be correct by DNA sequencing.The COS-7 cells transfected with pEGFP-N3-GAP-43 expressed GAP-43 protein efficiently.Conclusions: The cloning of rat GAP-43 gene and the construction of its eukaryotic expression vector were successful,which lays the foundation for further investigating the role of GAP-43 in development of oligodendrocytes.

       

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