| 小鼠GM-CSF基因的扩增及在HEK293T细胞中的分泌表达 |
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| 引用本文： | 李楠,张峰,仲飞.小鼠GM-CSF基因的扩增及在HEK293T细胞中的分泌表达[J].河北农业大学学报,2012,35(1):75-78. |
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| 作者姓名： | 李楠 张峰 仲飞 |
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| 作者单位： | 河北农业大学动物科技学院,河北保定,071001 |
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| 基金项目： | 国家自然科学基金(30771586);河北省自然科学基金(C2008000244) |
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| 摘 要： | 粒细胞-巨噬细胞集落刺激因子(GM-CSF)是机体免疫系统的重要细胞因子,具有生物佐剂作用,为研究GM-CSF的生物佐剂作用,本试验通过RT-PCR方法从小鼠脾脏细胞中扩增小鼠GM-CSF的cDNA,并将其插入到pcDNA3.1质粒中,构建成GM-CSF真核表达载体pcDNA3.1-GMCSF,并在真核细胞进行了瞬时表达。结果表明,本研究扩增的小鼠GM-CSF基因序列与GenBank序列完全一致,表达载体经脂质体介导转染HEK293T细胞,表达产物经western-blot检测,证明GM-CSF能够在HEK293T细胞中进行分泌表达。这为今后研究GM-CSF在动物疫苗,特别是DNA疫苗中的生物佐剂作用创造了必要的条件。
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| 关 键 词： | 小鼠GM-CSF 基因扩增 表达 |
Amplification of mouse GM-CSF gene and its expression in HEK293T cells |
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| LI Nan
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ZHANG Feng
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ZHONG Fei.Amplification of mouse GM-CSF gene and its expression in HEK293T cells[J].Journal of Agricultural University of Hebei,2012,35(1):75-78. |
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| Authors: | LI Nan ZHANG Feng ZHONG Fei |
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| Institution: | (College of Animal Science and Technology,Agricultural University of Hebei,Baoding 071000,China) |
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| Abstract: | Granulocyte and macrophage colony-stimulating factor(GM-CSF) is an important cytokine in animal immune system and is also a biological adjuvant.To express GM-CSF in a secretory manner in eukaryotic cells,the mouse GM-CSF cDNA was amplified with RT-PCR from mouse spleen and was inserted into pcDNA3.1A expression vector to construct its eukaryotic expression vector,pcDNA3.1-GMCSF.The transient expression of GM-CSF was performed in HEK293T cells transfected via liposome mediation.The results showed that the amplified GM-CSF gene sequence was consistent with that published in GenBank.The recombinant mouse GM-CSF was detected by Western-blot in the culture medium of the transfected HEK293T cells,which indicates that the GM-CSF gene could be expressed and secreted in the cells.Those results provide the necessary conditions for further study on the GM-CSF adjuvant functions in animal vaccine,especially in animal DNA vaccine. |
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| Keywords: | mouse GM-CSF gene amplification expression |
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