猪圆环病毒I型ORF2蛋白序列与其核定位功能的相关性

Relationship between PCV1-ORF2 fusion protein sequence and its nuclear localization

软件分析PCV1-ORF2的核苷酸序列发现其N端的氨基酸序列中包含一段核定位序列,将具有核定位序列特征的区域删除,构建缺失核定位序列的PCV1-ORF2基因与绿色荧光蛋白的真核融合表达载体,同时构建PCV1-ORF2全基因与绿色荧光蛋白的真核融合表达载体,分别转染Hela细胞,可观察到后者绿色荧光聚集在细胞核区域;而前者转染Hela细胞结果显示核定位现象消失,从而可以推断这一区域是CPV1-ORF2蛋白核定位的功能区。

After the analysis of the amino acid sequence of PCV1-ORF2, a domain of the amino acid sequence of the protein that has the character of nuclear localization sequences was found, deletion of this domain from PCV1-ORF2, the remainder domain was subcloned and fused to Enhanced Freen Fluorescence Protein（EGFP）. Then the fusion gene construct was transfected to Hela cells. The green fluorescence was observed around the whole cytosol. Meanwhile , a fusion gene construct which can express the fusion protein of EGFP and PCV1-ORF2 in eukaryotic cells was transfected into Hela cells, too. The green fluorescence could be seen to locate mainly in the nuclei.This means that the basic amino acids region near the N end is very important for PCV1-ORF2 fusion protein nuclear localization.