中文摘要：

用Trizol试剂提取红莲型水稻杂种F1幼穗的总RNA，分离纯化mRNA后反转录并合成双链cDNA．经Sephrose CL-2B凝胶过滤柱层析后，收集大于400bp以上的双链cDNA片段，将其连接到pTRG载体上，转化到XL1-Blue MRF’Kan感受态细胞后构建成红莲水稻幼穗细菌双杂交文库．经检测计算该原始文库容量为1．03×10^6 pfu／mL，对随机挑取的100个克隆进行PCR鉴定显示重组率接近100％而且大于800bp的插入片段达到97％．扩增后文库的容量达到1．05×10^10pfu／mL．

英文摘要：

The total RNA was extracted from the Young Panicle of F1 hybrid rice by Trizol reagent. The mRNA was purified from the total RNA by Poly-A Tract mRNA Isolation Kit and the double-stranded （ds） cDNA was synthesized after reverse transcription. The fragments longer than 400 bp in length were fractionated by Sephrose CL-2B Column, and then ligated into the pTRG vertor. The ligation product was transformed into XL1-Blue MRF Kan Library Pack Competent Cells to construct the bacterial two-hybrid system library. Analysis showed that the primary library contained 1.03 ×10^6 pfu/mL, and approximately 100% of the clones were positive recombinants and about 97 % of the cDNA fragments inserted were longer than 800 bp in length. The capacity of the amplified library was 1.05 ×10^10 pfu/mL.