华东葡萄白河-35-1株系抗霜霉病基因cDNA文库构建及EST分析

以中国原产抗霜霉病葡萄野生种华东葡萄（Vitis pseudoreticulata）白河-35-1为材料，用SMARTTM技术构建葡萄霜霉菌（Plasmopara viticola）诱导的葡萄叶片cDNA文库。经检测，原始文库滴度0.92×106 pfu/mL，重组率97%，插入片段500～2 200 bp，扩增文库滴度为5×109 pfu/mL。随机挑取克隆5'端测序，获得254条有效EST序列（GenBank登录号：FG269201~FG269449和FG396006~FG396010）。经BlastX分析，有56.3% ESTs与GenBank中功能已知基因有较高同源性，14.6%ESTs与功能尚未确定的假设蛋白或未知蛋白同源性很高，20.9%ESTs在GenBank中无匹配的同源产物。

Construction of cDNA Library and Analysis of Expressed Sequence Tags of Chinese Wild Vitis pseudoreticulata Clone Baihe-35-1 Resistance to Downy Mildew

A cDNA library was constructed with Vitis pseudoreticulata clone Baihe-35-1 leaves inoculated by Plasmopara viticola using SMARTTM cDNA Library Construction Kit. The titer of primary library and amplified library was about 0.92×106 pfu/mL and 5×109 pfu/mL, respectively. In the primary library, 97% clones were recombinant and the length of insert cDNAs were from 500 to 2 200 bp. Two hundred and fifty-four ESTs (GenBank accession No. FG269201~FG269449 and FG396006~FG396010) were obtained by random sequencing of the cDNA library. BlastX search showed that 56.3% ESTs had higher homology with functional genes, 14.6% ESTs with hypothetical putative or unknown protein, and 20.9% ESTs had no-hits.