转基因小麦B73-6-1品系特异性定性 PCR检测方法的建立

中国生物工程杂志

2011, Vol. 31

Issue (10): 100-105

技术与方法

转基因小麦B73-6-1品系特异性定性 PCR检测方法的建立

霍楠, 张明辉, 刘营, 仇有文, 敖金霞, 曲波, 高学军

东北农业大学生命科学与生物技术研究中心, 农业部转基因生物产品成分监督检验测试中心哈尔滨 150036

Establishment of Event Specific Qualitative PCR for Detecting Genetically Modified Wheat B73-6-1

HUO Nan, ZHANG Ming-hui, LIU Ying, QIU You-wen, AO Jin-xia, QU Bo, GAO Xue-jun

Northeast Agricultural University, Life Science and Biotechnique Research Center, Testing and Inspection for GMO of Ministry of Agriculture of P.R.C., Haerbin 150036, China

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摘要：

以转基因小麦B73-6-1为研究对象,通过染色体步行技术,成功分离到B73-6-1上pAHC25质粒外源基因插入位点的3'端旁侧序列,其扩增片段覆盖了转化载体及转基因小麦基因组旁侧序列。同时根据旁侧序列设计引物,建立品系特异性定性PCR检测方法,以典型的转基因作物证明该方法检测B73-6-1具有高特异性。该方法特异性好、灵敏度高, 可快速、准确、高效地检测转基因小麦B73-6-1品种。

关键词： 转基因小麦; B73-6-1; 染色体步行; 品系特异性; PCR检测

Abstract:

By Chromosome walking PCR technique, the 3'flanking sequence of the exogenous gene in the pAHC25 plasmid of wheat ‘B73-6-1’ was successfully cloned. An event specific transgenic detection method for ‘B73-6-1’ was established by PCR with the specific primers based on the 3′border. The amplification fragment include part sequence of both the exogenous DNA and wheat genome. This assay was applied to detect typical genetically modified crops, with high specificity for B73-6-1, and the limit of detection was 0.1%. The results showed that the qualitative PCR assay was accurate, rapid and efficient for detection of B73-6-1.

Key words: Genetically modified wheat B73-6-1 Chromosome walking Event-specific PCR detection

收稿日期: 2011-05-31 出版日期: 2011-10-25

ZTFLH:

Q819

基金资助:

转基因重大专项(2008ZX08012-001)资助项目

通讯作者: 高学军 E-mail: gaoxj5390@sina.com

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引用本文:

霍楠, 张明辉, 刘营, 仇有文, 敖金霞, 曲波, 高学军. 转基因小麦B73-6-1品系特异性定性 PCR检测方法的建立[J]. 中国生物工程杂志, 2011, 31(10): 100-105.

HUO Nan, ZHANG Ming-hui, LIU Ying, QIU You-wen, AO Jin-xia, QU Bo, GAO Xue-jun. Establishment of Event Specific Qualitative PCR for Detecting Genetically Modified Wheat B73-6-1. China Biotechnology, 2011, 31(10): 100-105.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I10/100

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