Ma Yimian, Liu Luo, Zhu Chengguang, Sun Changhui, Xu Bo, Fang Jun, Tang Jiuyou, Luo Anding, Cao Shouyun, Li Gupo, Qian Qian, Xue Yongbiao, Chu Chengcai
State Key Laboratory of Plant Genomics and National Plant Gene Research Center (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
J Genet Genomics. 2009 May;36(5):267-76. doi: 10.1016/S1673-8527(08)60114-9.
About 25,000 rice T-DNA insertional mutant lines were generated using the vector pCAS04 which has both promoter-trapping and activation-tagging function. Southern blot analysis revealed that about 40% of these mutants were single copy integration and the average T-DNA insertion number was 2.28. By extensive phenotyping in the field, quite a number of agronomically important mutants were obtained. Histochemical GUS assay with 4,310 primary mutants revealed that the GUS-staining frequency was higher than that of the previous reports in various tissues and especially high in flowers. The T-DNA flanking sequences of some mutants were isolated and the T-DNA insertion sites were mapped to the rice genome. The flanking sequence analysis demonstrated the different integration pattern of the right border and left border into rice genome. Compared with Arabidopsis and poplar, it is much varied in the T-DNA border junctions in rice.
利用具有启动子捕获和激活标签功能的载体pCAS04构建了约25000个水稻T-DNA插入突变体库。Southern杂交分析表明，约40%的突变体为单拷贝整合，平均每个突变体的T-DNA插入数为2.28。通过田间大规模表型分析，获得了许多具有重要农艺性状的突变体。对4310个初级突变体进行组织化学GUS分析，结果显示GUS染色频率高于以往报道，在各组织中均有表达，尤其在花中表达量较高。分离了部分突变体的T-DNA侧翼序列，并将T-DNA插入位点定位到水稻基因组上。侧翼序列分析表明，T-DNA左右边界在水稻基因组中的整合模式不同。与拟南芥和杨树相比，水稻T-DNA边界连接的变化更大。
