| 端粒蛋白TRF1与肌动蛋白结合蛋白PFN2相互作用的鉴定 |
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| 引用本文： | 马航航,杨平勋,黄翠芬,黄君健.端粒蛋白TRF1与肌动蛋白结合蛋白PFN2相互作用的鉴定[J].生物技术通讯,2011,22(3):384-388. |
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| 作者姓名： | 马航航 杨平勋 黄翠芬 黄君健 |
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| 作者单位： | 军事医学科学院,生物工程研究所,北京,100850 |
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| 摘 要： | 目的:鉴定端粒蛋白TRF1和肌动蛋白结合蛋白PFN2是否存在相互作用,并且两者的相互作用是否与端粒在细胞核周的锚定有关。方法:将TRF1构建到myc标签载体,PFN2构建到GST标签载体,采用GST-pull down技术,验证两者是否存在相互作用;同时将TRF1构建到EGFP标签的绿色荧光载体,PFN2构建到RED标签的红色荧光载体,两者共转入细胞,利用荧光显微镜观察两者在细胞中的共定位情况。结果:GST-pull down证明TRF1与PFN2存在直接相互作用,两者在细胞中可以共定位。结论:TRF1与PFN2存在相互作用,且这种相互作用发生在细胞核周。
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| 关 键 词： | 端粒 端粒酶 端粒相互作用蛋白 TRF1 PFN2 |
Identification of the Interaction of Telomere Binding Factor TRF1 and Actin Monomer-Binding Protein PFN2 |
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| MA Hang-Hang,YANG Ping-Xun,HUANG Cui-Fen,HUANG Jun-Jian.Identification of the Interaction of Telomere Binding Factor TRF1 and Actin Monomer-Binding Protein PFN2[J].Letters in Biotechnology,2011,22(3):384-388. |
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| Authors: | MA Hang-Hang YANG Ping-Xun HUANG Cui-Fen HUANG Jun-Jian |
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| Institution: | Beijing Institute of Biotechnology,Beijing 100850,China |
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| Abstract: | Objective：To identify the interaction between telomere binding protein TRF1 and actin monomer-binding protein PFN2,and study whether the interaction is related to the telomere anchoring to the nuclear membrane.Methods：TRF1 was constructed to the myc-tagged vector while PFN2 was constructed to the GST-tagged vector.GST-pull down assay was performed to verify the interaction of the two proteins.TRF1 and PFN2 was cloned to the EGFP-tagged and Red-tagged vectors respectively,and then co-transfected to HepG2 cells.Co-localization of the two protein in the cells was determined using the method of cyto-immunofluorescence.Results：GST pull-down assay proved the direct interaction between TRF1 and PFN2.Cyto-immunofluorescence analysis further confirmed the co-localization of the two proteins in the cell.Conclusion：The interaction between TRF1 and PFN2 is correlated to the telomere anchoring to the nuclear membrane. |
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| Keywords: | TRF1 PFN2 |
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