检测及鉴定RoundupReady转基因大豆寡核苷酸芯片制备的研究.pdfVIP

业生物技术学报 Journal of Agricultural Biotechnology 2005,13 (4): 429~434 ·研究论文· 检测及鉴定Roundup Ready 转基因大豆寡核苷酸芯片的制备* 1 2 2 1 1 小丹 文思远 王升启 黄昆仑 罗云波 ** ( 1.中国农业大学食品科学与营养工程学院, 北京 100083; 2. 北京放射医学研究所,北京 100850) ： 、种属相关 、 摘要 通过对Roundup Ready 转基因大豆(简称RRS)的基因盒结构分析，筛选了包含筛选基因 基因、目的基因 交联结构基因、特异结构基因、内对照基因、其它阴性控制基因和阳性参考目的基因的特异探针，制备了检测及鉴定Roundup Ready 转基因大豆的寡核苷酸芯片。实验表明，该芯片探针特异性好，可在一张芯片上完成对Roundup Ready 转基因大豆结构 的鉴定；内对照基因灵敏度高，检测极限为0.1 ng (DNA) ，从而消除假阴性概率；同常用的凝胶电泳检测方法相比，芯片杂交方 法灵敏度(RRS 为0.5%)优于凝胶电泳检测(RRS 为 1%)。 关键词: 转基因生物；寡核苷酸芯片；Roundup Ready 转基因大豆；检测及鉴定 Preparation of Oligonucleotide Microarray for Detecting and Identifying Roundup Ready Soybean XU Xiao-Dan 1 WEN Si-Yuan2 WANG Sheng-Qi 2 HUANG Kun-Lun 1 LUO Yun-Bou1 To detect and identify transgenic soybean, Roundup Ready soybean(RRS), a microarray consisting of screening probe, species reference probe, specific probe, construct specific probe, event specific probe, internal control probe, negative control probe and reference specific probe was constructed by analyzing transgentic structure of RRS. All the probes were evaluated as specific. De- tecting and identifying RRS structure could be completed in one microarray. The internal control with high sensitivity (0.1 ng DNA) could eliminate possibility of false negative. The sensitivity of oligonucleotide microarray (RRS 0.5%) was higher than that of the electrophoresis method (RRS 1%). genetically m