绵毛优若藜冷胁迫均一化全长cDNA文库的构建

Construction of cold-inducible normalized full-length cDNA library of Ceratoides lanata

以绵毛优若藜叶片和嫩茎为材料，对盆栽的绵毛优若藜小苗进行梯度低温胁迫处理后，采用SMART（switching mechanism at 5’end of RNA transcript）与DSN（duplex-specific nuclease）均一化相结合技术构建绵毛优若藜冷胁迫均一化全长cDNA文库。经测定，原始文库的库容量为1．4×10^6pfu。PCR检测结果显示：插入片段的长度在0．5～3kb之间，平均大于1kb，表明文库构建效果较好。该文库包含有大量的未知基因，有待进一步的发掘和研究。

Taking the leaves and young stems as material, a normalized full-length eDNA library of Ceratoides lanata was constructed. The seedlings of potted Ceratoides lanata were treated by gradient cold stress, normalized fulllength eDNA library was successfully constructed by DSN （duplex-specific nulease）-normalization combined with SMARTTM （switching mechanism at 5＇end of RNA transcript） technique. The results show that the capacity of the primary library was 1.4×10^6pfu. The PCR results show that the insertion size was between 0. 5 kb and 3. 0 kb, averagely about 1.0 kb, suggesting that the eDNA library was well normalized and had been established successfully. The analytic result suggested that this eDNA library contains large new genes, which need further researches and function- al analysis.