摘要
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。
With the aim to investigate the expression pattern of genes at different ripening stages so as to select the anti-insect,disease-resistant and high-yield genes,the veraison skin（50% colored） of Vitis amurensis was used as experimental material to construct the cDNA library.The pDNR-LIB was used as the vector and the recombinant plasmid was transformed into E.coli DH5α by the electroporation according to the CreatorTM SMARTTM Library construction Kit.The fragment size inserted was 0.5 to 2.0 kb and the average insert size was 0.80 kb.710 unigenes obtained were clustered into 97 contigs and 613 singlets.The result of the BlastX analysis indicated that there were 528 ESTs with known or unknown functions,182 ESTs with no significant similarity matching any protein or DNA sequence in the database.Some genes involved in the process of anthocyanin synthesis and defense/disease-resistant genes were detected in this library,such as glutathione S-transferase,zinc finger protein,chalcone synthase,and anthocyanidin 3-O-glucosyltransferase,which inferred that these genes were expressed actively at the veraison stage.
出处
《果树学报》
CAS
CSCD
北大核心
2011年第4期597-602,共6页
Journal of Fruit Science
基金
现代农业产业技术体系建设专项资金(nycytx-30)
