中文摘要：

噬菌体展示是研究蛋白质相互作用的重要手段，为深入研究红莲型水稻不育和育性恢复的分子机理，以红莲型水稻杂种F1花药为材料，分离纯化mRNA，经反转录合成双链eDNA，在双链eDNA末端加上定向EcoRⅠ／Hind Ⅲ接头，再用EcoRⅠ和HindⅢ消化接头，形成两端分别带有EcoRⅠ和Hind Ⅲ粘性末端的双链eDNA。经Mini Column纯化后，收集300 bp以上的双链eDNA片段，将其连接到带有EcoRⅠ和Hind Ⅲ末端的T7 Select 10-3b载体上，经体外包装后，以BL T5403为受体菌构建了红莲水稻杂种F1花药的噬菌体展示文库。经测定显示，该噬菌体展示文库容量为1．03×10^6pfu／mL，重组率为100％，扩增后文库滴度为2．14×10他pfu／mL^6对随机挑取的100个噬菌斑进行PCR鉴定，97％的插入片段大干300 bp。

英文摘要：

Phage display is a powerful method to study protein-protein interactions. In order to study the molecular mechanism of cytoplasmic male sterility and fertility restoration in Honglian rice, the mRNA was isolated with PolyA Tract mRNA Isolation Kit from the anther of F1 hybrid rice and the double strand （ds） cDNA was synthesized by reverse transcription. Then the directional EcoR Ⅰ IHind Ⅲ linkers were ligated into the ends of ds cDNA and the ds cDNA was further digested with EcoR Ⅰ and Hind Ⅲ, which resulted in ds cDNA with EcoR Ⅰ and Hind Ⅲ ends. The digested ds cDNA flagments longer than 300 bp in length were fractionated with Mini Column, then ligated into the T7 Select 10-3b vertor with EcoR Ⅰ and HindⅢends. After packaging in vitro, the T7 Select 10-3b vertor was tansformed into BL T5403 to construct the T7 phage display library. Analysis showed that the library contained 1.03×106 clones per microliter, and approximately 100% of the clones in library was recombinant. The titer of the amplied library was 2.14×10^12 pfu/mL, and the insert length of the recombinahts over 300 bp was about 97%.