转基因水稻吉生粳2号的外源基因旁侧序列分离及事件特异性PCR检测方法  

Identification of the T-DNA Flanking Sequences and Event-Specific PCR Detection of Transgenic Rice‘Jishengjing 2'

作　　者：金永梅[1] 马瑞[1] 于志晶[1] 林秀峰[1] 

机构地区：[1]吉林省农业科学院农业生物技术研究所,长春130033

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)

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＇.

关 键 词：转基因水稻 旁侧序列 事件特异性检测 

分 类 号：S511.035.3[农业科学—作物学]