| 摘要: |
| 本试验针对已商品化的转基因大豆、玉米和水稻的主要外源基因Cry1A(B)基因、BAR基因、CP4-EPSPS基因和PAT基因及共有的内标准基因RBCL基因设计了10对引物,对从超市购买的进口大豆、玉米和水稻的11种深加工制品(未标注是否含有转基因成分)进行了五重巢式PCR定性检测。第1轮普通五重PCR检测,11个待测样品均未出现任何转基因成分, 其灵敏度为 0.5% ;第2轮将普通五重PCR扩增产物进行五重巢式PCR检测,结果在卵磷脂、大豆蛋白质粉、巧克力饮品和婴儿米粉中扩增出RBCL基因,玉米淀粉和玉米泥中扩增出RBCL基因、Cry1A(B)基因和PAT基因;玉米蛋白粉扩增出RBCL基因、Cry1A(B)基因和BAR基因,营养麦片扩增出内参RBCL基因和Cry1A(B)基因,在大豆精炼油、色拉油和玉米油中未检测出上述2种基因和另外3种外源基因CP4-EPSPS、BAR和PAT,其检测灵敏度为 0.005%。结果提示,五重巢式 PCR 检测方法适用于转基因大豆、玉米和水稻深加工产品的定性检测。 |
| 关键词: 转基因 大豆 玉米 水稻 深加工产品 五重巢式 PCR |
| DOI：10.11841/j.issn.1007-4333.2010.02.017 |
| 投稿时间：2009-03-10 |
| 基金项目:东北农业大学创新团队资助项目(CXT004-3-2); 转基因重大专项(2008ZX08012-001); 黑龙江省青年科学基金资助项目(QC2009C49)第一作者:敖金霞,助理研究员,博士,主要从事转基因作物基因检测技术研究,E-mail: aojinxia@yahoo.com.cn通讯作者:高学军,教授,主要从事生物化学与分子生物学研究,E-mail: gaoxj5390@sina.com.cn |
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| Detection of transgenic soybean, maize and rice in highly processed products by multiple nested PCR |
|
AO Jin-xia, GAO Xue-jun, YU Yan-bo, QU Bo, LI Qing-zhang
|
| (Life Science and Biotechnique Research Center of Northeast Agricultural University/Testing and Inspecting Center for GMO, Harbin 150030, China) |
| Abstract: |
| Ten pairs of primers were designed for alien foreign genes, Cry1A(B) gene, BAR gene, PAT gene and common reference RBCL gene inserted in commercialized transgenic soybean, maize and rice to detect eleven highly processed products samples from supermarket by the multiplex nested PCR. The first common multiple PCR could not detect the insert signals in eleven processed products, and the sensitivity is 0.5％. However the amplification by the second multiplex nested PCR using the result of common multiple PCR for templates, which could simultaneously detect RBCL and CP4-EPSPS genes in soya lecithin, soya protein powder, chocolate beverage and infant rice cereal; RBCL ,Cry1A(B) and PAT genes in maize starch and maize jam; RBCL ,Cry1A(B) and BAR genes in maize protein powder; RBCL and Cry1A(B) genes in oatmeal, and not detect the said two genes and the three foreign CP4-EPSPS, BAR, PAT genes in soybean refine oil, soybean salad oil and maize oil. The sensitivity of the multiplex nested PCR is 0.005％. The results indicate that multiplex nested PCR is suited to qualitative detection for transgenic soybean, maize and rice in highly processed products. |
| Key words: transgenic soybean maize rice highly processed products multiple nested PCR |
