基于焦磷酸测序技术建立基因编辑水稻检测方法  

Pyrosequencing-based Detection Method of Gene Edited Rice

作　　者：解美霞 王燕 赵新[1] 高越 刘双 陈锐[1] 兰青阔[1] 徐晓辉[4] 曹际娟 王永[1] XIE Meixia;WANG Yan;ZHAO Xin;GAO Yue;LIU Shuang;CHEN Rui;LAN Qingkuo;XU Xiaohui;CAO Jijuan;WANG Yong(Institute of Biotechnology,Tianjin Academy of Agricultural Sciences,Tianjin 300381,China;College of Agronomy,Hebei Agricultural University,Hebei Baoding 071001,China;College of Food Science and Technology,Hebei Agricultural University,Hebei Baoding 071001,China;Shandong Academy of Agricultural Sciences,Jinan 250100,China;Dalian Minzu University,Liaoning Dalian 116600,China)

机构地区：[1]天津市农业科学院生物技术研究所,天津300381 [2]河北农业大学农学院,河北保定071001 [3]河北农业大学食品科技学院,河北保定071001 [4]山东省农业科学院,济南250100 [5]大连民族大学,辽宁大连116600

出　　处：《生物技术进展》2020年第6期668-673,共6页Current Biotechnology

基　　金：国家转基因新品种培育重大专项(2019ZX08010-002;2019ZX08015-002)。

摘　　要：基因编辑技术发展迅速,但对应的检测方法较少。为寻找创建基因编辑作物适用的检测方法,以PL 3基因编辑水稻编辑位点为靶标,有效设计了焦磷酸测序的扩增引物及测序引物,并进行有效性检测,分别利用Sequence to Analyze等程序以及SNP和AQ两种模式完成了对PL 3基因的定性和定量检测试验,建立了PL 3基因编辑水稻编辑位点焦磷酸测序检测方法。结果表明,基于焦磷酸测序技术可以通过检测编辑位点从而将基因编辑型水稻与野生型水稻进行区分。与常规的转基因检测方法相比,该检测方法具有较好的准确性、高效性及高灵敏度等优点,在基因编辑型水稻编辑位点定性和定量检测分析方面具有很好的应用前景。Gene editing technology is developing rapidly,but there are few related detection methods.In order to find a detection method for gene-edited crops,this study used editing sites of PL 3 gene-edited rice as the target,effectively designed pyrosequencing amplification primers and sequencing primers,and conducted effectiveness testing,respectively using Sequence to Analyze and other programs as well as the two models of SNP and AQ,the qualitative and quantitative detection test of PL 3 gene has been completed,and the pyrosequencing detection method on editing site of PL 3 gene-edited rice has been established.The results showed that the editing type rice could be distinguished from the wild type rice by detecting the editing sites based on pyrosequencing.Compared with the conventional methods,the method has the advantages of better accuracy,high efficiency and high sensitivity.It has a good application prospect in qualitative and quantitative detection of editing sites at the gene editing rice.

关 键 词：焦磷酸测序 PL 3基因 基因编辑 水稻 

分 类 号：Q943.2[生物学—植物学] S511[农业科学—作物学]