山葡萄(双丰)转色期果皮cDNA文库构建和分析Construction and analysis of cDNA library from Vitis amurensis grapevine skins at veraison
李波;杨成君;刘海峰;李凤;段长青;王军;
摘要(Abstract):
为了研究山葡萄不同成熟期基因表达的规律,从而筛选抗病、抗虫和高产的优质基因,以转色期(50%着色)山葡萄果皮为材料,采用Clontech公司的cDNA文库构建试剂盒以pDNR-LIB为载体,采用电穿孔法将重组质粒转化到DH5α中,构建了山葡萄果皮cDNA文库。文库插入片段大小为0.5~2.0 kb,平均约为0.80 kb。随机挑取1 009个克隆进行测序,共拼接出710个Unigenes,其中包括97个重叠群(Contigs),613个单拷贝(Singlets)。经BlastX分析共获得已知功能和未知功能基因528个,无序列相似新基因182个。文库中涉及了花色苷合成途径和抗病防御的相关基因,如谷胱甘肽-S-转移酶、锌指蛋白、查尔酮合成酶和花青素3-O-葡萄糖苷转移酶等,可见这些基因在转色期时表达比较活跃。
关键词(KeyWords): 山葡萄;cDNA文库;转色期;EST测序
基金项目(Foundation): 现代农业产业技术体系建设专项资金(nycytx-30)
作者(Author): 李波;杨成君;刘海峰;李凤;段长青;王军;
Email:
DOI: 10.13925/j.cnki.gsxb.2011.04.019
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