前列腺癌上调表达基因BRP44的蛋白细胞定位及功能预测研究

Cellular localization of BRP44 up-expressed in prostate cancer and prediction of gene function

目的了解GFP-BRP44融合蛋白的细胞定位,初步预测BRP44的功能。方法构建BRP44与GFP的融合表达真核载体,脂质体介导法转染高表达该基因的前列腺癌细胞株LNCaP,激光共聚焦显微镜观察融合蛋白的细胞定位。生物信息学方法对基因BRP44进行序列分析及功能预测。结果成功构建GFP-BRP44融合蛋白的真核表达载体,pEG-FP-BRP44融和蛋白在LNCaP细胞核及细胞质均有表达。与单纯GFP相比,融合蛋白在细胞质的表达相比细胞核更强。功能预测提示基因BRP44可能是一个受oct-1等转录因子调控,主要在细胞线粒体发挥基因调节作用的可溶性蛋白。结论BRP44蛋白可能主要在细胞质发挥作用。

Objective To explore the subcellular localization of GFP-BRP44 in LNCaP cells and predict the gene functions of BRP44. Methods The plasmid with fusion gene of BRP44 and green fluorescent protein （GFP） was constructed and transfected into LNCaP cells by liposome. The subcellular localization of GFP- BRP44 and pure GFP was observed by laser scannning confocal microscope. The gene function was predicted by analyzing the sequence and motif of BRP44 with bioinformatics. Results Some useful information predicted about gene function of BRP44 was gotten. The fusion protein of GFP-BRP44 expressed in both nuclei and cytoplasm of LNCaP cells. However, compared with pure GFP, the expression of GFP-BRP44 in cytoplasm was stronger than in nuclei. Predicted information hinted that BRP44 may be a regulatory factor regulating some gene expression in cellular mitochondria. Conclusion The protein of BRP44 mainly located in cytoplasm of LNCaP cells.