端粒蛋白TRF1与肌动蛋白结合蛋白PFN2相互作用的鉴定  

Identification of the Interaction of Telomere Binding Factor TRF1 and Actin Monomer-Binding Protein PFN2

作　　者：马航航[1] 杨平勋[1] 黄翠芬[1] 黄君健[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850

出　　处：《生物技术通讯》2011年第3期384-388,共5页Letters in Biotechnology

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。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.

关 键 词：端粒 端粒酶 端粒相互作用蛋白 TRF1 PFN2 

分 类 号：Q24[生物学—细胞生物学] Q78