葡萄PRC1-like核心组分基因克隆及序列分析

Cloning and Sequence Analysis of PRC1-like Core Components Genes in Grapevine

克隆葡萄PRC1-like核心组分基因并预测其功能。在NCBI中分别查找与拟南芥PRC1核心组分基因同源性最高的葡萄序列,采用CTAB法提取葡萄品种‘赤霞珠’(Cabernet sauvignon)嫩叶中总RNA,运用RT-PCR技术进行克隆,得到葡萄PRC1-like核心组分基因的CDS序列,并通过各种在线软件对葡萄PRC1-like核心组分基因及其编码的蛋白质进行生物信息学分析。获得4个葡萄PRC1-like核心基因,这些基因CDS全长分别为1 614 bp、1 362 bp、1 293 bp和1 275 bp,分别编码568个、453个、430个和424个氨基酸,预测其蛋白质的相对分子量分别为64.17 kD、50.69 kD、47.99 kD和47.71 kD,推测这些蛋白均为亲水性不稳定蛋白,与已知拟南芥PRC1相应的核心蛋白具有高度的同源性,亚细胞定位预测结果表明这些蛋白主要存在于细胞核中。推测葡萄PRC1-like在葡萄发育中起着重要作用,这为进一步阐明葡萄PRC1-like的功能及作用机制提供了理论依据。

The full-length CDS sequences of PRC1-like core components genes were cloned from grapevine and their functions were predicted and analyzed. To identify genes with homology to Arabidopsis PRC1 genes in grapevine genome, we performed BLAST searches with Arabidopsis PRC1 nucleotides and proteins sequences as a query as well. The total RNA was extracted from young leaves of‘Cabernet sauvignon’by CTAB method. The CDS sequences of grapevine PRC1-like core components genes were amplified by RT-PCR. The grapevine PRC1-like core component gene nucleotides and amino acid sequences were analyzed by various online bioinformatics softwares. The four genes of grapevine PRC1-like core components were obtained. And their full-length CDS are 1 614 bp, 1 362 bp, 1 293 bp and 1 275 bp, respectively, and they encode 537, 453, 430 and424 amino acids, respectively. The relative molecular weight of coded protein of grapevine PRC1-like core components genes are 60.58 kD, 50.69 kD, 47.99 kD and 47.71 kD, respectively. All of them are presumed to be hydrophilic labile proteins, which are highly homologous to the known core proteins of Arabidopsis thaliana PRC1. The prediction results of subcellular localizations showed that grapevine PRC1-like proteins are nuclear proteins. It predicted that grapevine PRC1-like might play an important role in the development of grapevine,which lays an information foundation for further study of the function and mechanism of grapevine PRC1-like.