BACKGROUND Cymbidium faberi is one of the oldest cultivars of oriental orchids, with an elegant flower fragrance. In order to investigate the molecular mechanism and the functions of related proteins in the methyl jasmonate (MeJA) signaling pathway, one of the main components of flower fragrance in C. faberi, yeast one- and two-hybrid three-frame cDNA libraries were constructed. RESULTS In this study, a modified cDNA library used for yeast one- and two-hybrid screening was successfully constructed, with a recombinant efficiency of 95%. The lengths of inserted fragments ranged from 750~3000 bp, and the library capacity reached 6 × 109 CFU/ μg of cDNA insert, which was suitable for the requirements of subsequent screening. Finally, a homologous protein related with pathogenesis was screened out by the bait vector of CfbHLH36, which may participate in the MeJA signaling pathway. CONCLUSION The yeast one- and two-hybrid library of C. faberi provides large amounts of useful information for the functional genomics research in C. faberi, and this method could also be applied to other plants to screen DNA-protein and protein-protein interactions.

中文翻译：

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适用于酵母一杂交和二杂交检测的大花C兰高质量cDNA文库的构建和表征。

背景技术大花ym兰是东方兰花中最古老的品种之一，具有优雅的花香。为了研究茉莉酸甲酯（MeJA）信号通路中相关蛋白的分子机制和功能，构建了梭状芽孢杆菌中花香的主要成分之一，酵母一，二杂交三框架cDNA文库。 。结果在本研究中，成功​​地构建了用于酵母一杂交和二杂交筛选的修饰的cDNA文库，重组效率为95％。插入片段的长度在750〜3000 bp之间，文库容量达到6×109 CFU /μgcDNA插入片段，适合后续筛选。最后，通过诱饵载体CfbHLH36筛选出与发病相关的同源蛋白，可能参与MeJA信号通路。结论Fab。faberi的酵母一和二杂交文库为Fab。faberi的功能基因组学研究提供了大量有用的信息，该方法还可用于其他植物以筛选DNA-蛋白质和蛋白质-蛋白质相互作用。