中文摘要：

目的：探讨A型核纤层蛋白前体（prelamin A）在细胞内堆积造成细胞早老的机理,筛选了prelamin A相互作用蛋白并研究其在早老细胞中的表达情况。方法：以prelamin A的C末端区域为诱饵蛋白,采用酵母双杂交方法从人骨骼肌cDNA文库中筛选prelamin A相互作用蛋白。构建了prelamin A识别因子（Narf）与绿色荧光蛋白融合表达载体pEGFP-Narf,与红色荧光蛋白-prelamin A融合表达质粒pDsRed-PLA共转染HEK293细胞,激光共聚焦显微观察共定位情况。Western blotting检测Narf在衰老表型HEK293PLA细胞的表达情况。结果：筛选得到包括Narf在内的7个候选相互作用蛋白。Narf与prelamin A能相互作用并共定位于核纤层,在prelamin A过表达的HEK293PLA细胞中Narf表达没有升高。结论：Narf在细胞内与prelamin A相互作用,且表达量不受后者影响。

英文摘要：

Objective：In order to reveal the mechanism of cell aging resulted from accumulation of prelamin A,proteins interacted with prelamin A were screened from human skeletal muscle library and the expression of nuclear prelamin A recognition factor（Narf） was analysized.Method：A bait plasmid pGBKT7-PLA-C was constructed containing C-teminal amino acids 389-664 of prelamin A and cotransformed into AH109 yeast competent cells with human skeletal muscle library.Narf gene was inserted into eukaryotic vector pEGFP generating the recombinant vector pEGFP-Narf,which was co-transfected into HEK293 cells with pDsRed-PLA and visualized by laser confocal microscopy.Expression of Narf in HEK293PLA cells was identifed by Western blotting.Result：Seven candidate proteins that interacted with prelamin A were identified by yeast two-hybrid screening including Narf.Colocalization assay showed that Narf and prelamin A were co-located around nucleus of HEK293 cells.The expression of Narf was not increased in the HEK293PLA cells,in which prelamin A was over expressed.Conclusion：Narf was able to interact with prelamin A and the latter can＇t affect Narf＇s expression.