摘要
利用染色体步移方法从转基因水稻吉生粳2号基因组中分离了外源基因T-DNA整合左边界旁侧序列,长度为297 bp。通过对左边界旁侧序列的同源性比对分析定位了外源基因T-DNA在吉生粳2号基因组中的整合位点,为水稻基因组第1号染色体的第41 607 441 bp处(Gen Bank sequence ID:NC_008394.4)。依据整合位点右侧设计的1条引物和TDNA右边界设计的3条引物分别进行PCR扩增分离到了右边界旁侧序列。根据左边界旁侧序列和T-DNA整合位点,建立了吉生粳2号事件特异性定性PCR检测方法,该方法为转基因水稻吉生粳2号的身份识别提供了有效手段。
Using genome walking PCR, left flanking sequence of the T-DNA integration site on rice genome was am- plified. T-DNA integration site on rice genome was determined by nucleotide blast using left flanking sequence as query. The results showed that T-DNA was inserted into the position 41 607 441 bp of chromosome 1 （GenBank sequence ID： NC_008394.4）. Right flanking sequence of the T-DNA integration site was also amplified by using T- DNA right border specific primers and right primer of insertion site. Based on the left flanking sequence and insertion site, the T-DNA left border specific primer and insertion site specific primers on ‘Jishengjing 2＇ genome were designed. Using the primers, the event-specific detection PCR for ‘Jishengjing 2＇ was established. It will provide the effective method for identification of ‘Jishengjing 2＇.
出处
《东北农业科学》
2016年第1期14-19,共6页
Journal of Northeast Agricultural Sciences
基金
吉林省农业科技创新工程项目(吉教厅[2012]1072号)
吉林省留学人员科技创新创业项目(RL201322)
吉林省科技发展计划项目(20140204014NY)
农业部转基因生物新品种培育科技重大专项(2014ZX08001-001-009)
关键词
转基因水稻
旁侧序列
事件特异性检测
Transgenic rice
Flanking sequences
Event-specific detection
