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

作　　者：明付焕[1] 张党权[1] 宋志丹[1] 谷振军[1] 祝全东[1] 韩欣[1] 郭林林[1] 

机构地区：[1]中南林业科技大学国家林业局栽培与育种重点实验室,湖南长沙410004

出　　处：《中南林业科技大学学报》2011年第7期169-173,共5页Journal of Central South University of Forestry & Technology

基　　金：国家自然科学基金项目(30700643;30972343);中南林业科技大学研究生科技创新基金资助项目(2008sx09);湖南省研究生科技创新基金(CX2010B323)

摘　　要：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.

关 键 词：绵毛优若藜 全长CDNA文库 均一化 冷胁迫 抗逆基因 

分 类 号：Q785[生物学—分子生物学]