| 本文已被：浏览 1097次 下载 612次 |
码上扫一扫！ |
| 转基因大豆OsDREB3品系特异性定性PCR检测方法的建立 |
|
刘营1,2, 张明辉1,2, 霍楠1,2, 仇有文1,2, 敖金霞1,2, 高学军1,2
|
|
|
| (1.东北农业大学 生命科学与生物技术研究中心, 哈尔滨 150036;2.农业部转基因生物产品成分监督检验测试中心(哈尔滨), 哈尔滨 150036) |
|
| 摘要: |
| 为建立转基因大豆OsDREB3品系特异性定性检测方法,以转基因大豆OsDREB3为研究对象,通过染色体步行方法,成功获得转基因大豆OsDREB3上pCAMBIA1301质粒外源基因插入位点的5'端旁侧序列,其扩增片段覆盖了转化载体及转基因大豆基因组旁侧序列,扩增片段大小为344 bp。同时根据旁侧序列设计引物,建立OsDREB3品系特异性定性PCR方法,以典型的转基因作物证明该方法检测转基因大豆OsDREB3具有高特异性,灵敏度为0.1%。 |
| 关键词: 转基因大豆 OsDREB3 染色体步行 品系特异性 PCR检测 |
| DOI：10.11841/j.issn.1007-4333.2012.04.006 |
| 投稿时间：2012-02-16 |
| 基金项目:转基因重大专项资助项目(2011ZX08004-002) |
|
| Establishment of event specific qualitative PCR for detecting transgenic soybean OsDREB3 |
|
LIU Ying1,2, ZHANG Ming-hui1,2, HUO Nan1,2, QIU You-wen1,2, AO Jin-xia1,2, GAO Xue-jun1,2
|
| (1.Life Science and Biotechnique Research Center, Northeast Agricultural University, Harbin 150036, China;2.Supervision and Test Center(Harbin) for Molecular Characteristics of Genetically Modified Plantsof Ministry of Agriculture, Harbin 150036, China) |
| Abstract: |
| By chromosome walking PCR technique,the 5'flanking sequences of the exogenous integration in the genome of in the pCAMBIA1301 plasmid of transgenic soybean OsDREB3 were cloned,and sequenced with the specific nested primers based on the border.According to the flanking sequence,the event specific primers were designed to amplify the fragment,which could span the exogenous DNA and carnation genome,and length was 344 bp.The event specific qualitative PCR detection method for transgenic soybean OsDREB3 was established.This assay was applied to detect typical genetically transgenic soybean OsDREB3,with high specificity,and the limit of detection exhibited 0.1%.The results showed that the qualitative PCR assay was accurate,rapid and efficient for detection of transgenic soybean OsDREB3. |
| Key words: transgenic soybean OsDREB3 chromosome walking event specificity PCR detection |
