‘津田’芜菁消减cDNA文库的构建及分析

Construction and Analysis of ‘Tsuda’ Turnip Subtracted cDNA Library

以‘津田’芜菁红色块根和白色块根为材料，利用SSH技术成功构建了消减cDNA文库并富集相关基因片段。从消减文库中筛选到960个阳性克隆，PCR鉴定插入片段长度主要分布于300～800bp之间。克隆测序后与GenBank中的同源序列进行比较，特异基因片段为302个，其中99个与数据库中的序列具有相似区域且功能已知，与数据库中序列没有显著同源性及功能未知的序列为203条。

A substracted cDNA library was constructed by using SSH technique to enrich related genes. cDNA prepared from red root peel of ‘Tsuda＇ tumip was used as tester, and that from white ones was used as driver. 960 positive clones were got from the library. The inserted size was between 300 and 800 base pair as it was determined by PCR method. After sequencing and clustering, 302 unique genes were obtained. By running blastn program of NCBI, 99 ESTs were found to have the homologous genes of the database with known function, and 203 ESTs without remarkable homology or with unknown function.