关键词: AtZW10, 生物信息学分析, 载体, 融合蛋白, 亚细胞定位

Abstract: The bioinformatics analysis of AtZW10 protein was performed by using different approaches. The results showed that the Arabidopsis thaliana AtZW10 protein is much conserved during the evolution of high eukaryotes. In AtZW0 protein, there is a similar conserved region to the centromere/kinetochorebinding protein ZW10. Moreover, the high similarity was found between AtZW0 and the Drosophila DmZW10 protein. AtZW10 was considered to be a hydrophilic protein by hydrophobicity analysis, and no obvious transmembrane domains. Prediction results also indicated that AtZW10 is likely to be located in the nucleus and cytoplasm. In order to further determine the subcellular localization of AtZW10, the full-length of AtZW10 gene was amplified by PCR using Arabidopsis cDNA as a template, and the binary vectors consisting of AtZW10 fused with the yellow (green) fluorescent protein (YFP/GFP) coding sequence were further constructed. The recombinant vectors were transformed into wild-type Arabidopsis protoplasts of mesophyll cells and Nicotiana benthamiana epidermal cells, respectively. Subsequently, the analyses on the distribution of the fusion proteins suggest that AtZW10 is an nucleoplasm positioning protein, and is mainly distributed in the nucleus, consistent with the conclusions of bioinformatics analysis. The molecular features and the subcellular localization of AtZW10 were analyzed in the present study, which might provide a basis for studying the function of AtZW10 gene in the future.

Key words: AtZW10, bioinformatic analysis, vector, fusion proteins, subcellular localization