基于KASP平台的转基因纯合体高通量检测方法的建立  

Establishment of a High-Throughput Homozygous Detection Method for Transgenic Maize Based on the KASP Platform

作　　者：孙铭勖 葛建镕[2] 朱维佳 王蕊[2] 赵怡锟 金肇阳 赵紫薇 朱少喜 赵久然[2] 王凤格[2] SUN Ming-xu;GE Jian-rong;ZHU Wei-jia;WANG Rui;ZHAO Yi-kun;JIN Zhao-yang;ZHAO Zi-wei;ZHU Shao-xi;ZHAO Jiu-ran;WANG Feng-ge(Faculty of Agronomy,Jilin Agricultural University,Changchun 130118;Maize Research Center,Beijing Academy of Agriculture&Forestry Sciences,Beijing 100097;Agricultural College,Yangzhou University,Yangzhou 225009;College of Advanced Agriculture and Ecological Environment,Heilongjiang University,Harbin 150008,China)

机构地区：[1]吉林农业大学农学院,长春130118 [2]北京市农林科学院玉米研究所/玉米DNA指纹及分子育种北京市重点实验室,北京100097 [3]扬州大学农学院,江苏扬州225009 [4]黑龙江大学现代农业与生态环境学院,哈尔滨150008

出　　处：《玉米科学》2024年第8期31-37,共7页Journal of Maize Sciences

基　　金：北京学者计划项目(BSP041);北京市农林科学院科技创新能力建设专项(KJCX20230301);北京市农林科学院财政追加专项(CZZJ202206)。

摘　　要：以转基因玉米DBN9501为材料,设计一套包含1条位于基因组的共用引物、两条分别位于外源插入序列与外源插入序列等位基因组序列的竞争性引物,能分别扩增出外源片段与基因组片段的纯合体检测引物,建立一种基于KASP平台的高通量转基因玉米纯合体检测方法。利用DBN9501BC1和15份其他转基因玉米样品进行引物评估、体系优化以及特异性验证。结果显示,所设计的引物既能特异性地对DBN9501进行成分检测,又能对DBN9501BC1代中的纯合阳性、杂合阳性以及阴性3种基因型进行准确鉴别。In this study,transgenic maize DBN9501 was used as the material.A set of primers was designed,in⁃cluding one common primer located in the genome and two competitive primers located in the exogenous insertion sequence and its allelic genome sequence,respectively.These primers can amplify the exogenous fragment and the genome fragment separately,a high-throughput method for detecting homozygous transgenic maize based on the KASP platform was established.Primers were evaluated,optimized,and validated for specificity using DBN9501BC1 and 15 other transgenic maize samples.The results showed that the designed primers could specifi⁃cally detect the composition of DBN9501 and accurately distinguish among the three genotypes:homozygous posi⁃tive,heterozygous positive,and negative in DBN9501BC1.

关 键 词：转基因玉米 纯合体检测 高通量 KASP平台 分子标记 

分 类 号：S513.035.3[农业科学—作物学]