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| 棉花中转基因成分多重PCR检测体系的建立 |
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陈贞1, 芦春斌2, 杨梦婕2, 马骏3, 白卫斌1, 吴希阳1
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| (1.暨南大学 理工学院,广州 510632;2.暨南大学 生殖免疫研究所,广州 510632;3.广东出入境检验检疫局 检验检疫技术中心,广州 510623) |
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| 摘要: |
| 以棉花内源基因sad1、报告基因GUS、外源抗虫基因Cry1Ab/Ac、筛选基因NPTⅡ、NOS终止子和CaMV35S启动子为检测对象,设计的6对引物能扩增长度合适的基因片段且避免了引物二聚体。通过优化PCR扩增体系中不同引物终浓度的配比以及退火温度对多重PCR检测的影响,建立了棉花转基因成分6重PCR检测体系。结果表明:采用本研究建立的棉籽基因组DNA提取方法,该6重PCR检测体系能够有效地从少至1颗棉籽的少量样品里检测出棉花中的转基因成分。 |
| 关键词: 棉花 转基因检测 多重PCR |
| DOI：10.11841/j.issn.1007-4333.2011.03.003 |
| 投稿时间：2010-09-13 |
| 基金项目:农业部转基因重大专项(2008ZX08012-005) |
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| Multiplex PCR for detection of genetically modified componentsin cotton seeds |
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CHEN Zhen1, LU Chun-bin2, YANG Meng-jie2, MA Jun3, BAI Wei-bin1, WU Xi-yang1
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| (1.College of Science and Engineering,Jinan University,Guangzhou 510632,China;2.Institute of Reproductive Immunology,Jinan University,Guangzhou 510632,China;3.Inspection and Quarantine Technology Center,Guangdong Entry-Exit Inspection andQuarantine Bureau,Guangzhou 510623,China) |
| Abstract: |
| To detect transgenic components in genetically modified cotton,endogenous gene sad1,reporter gene GUS,exogenous insect resistant gene Cry1Ab/Ac,screening gene NPTⅡ,NOS terminator and CaMV35S promoter were chosen for establishing a multiplex polymerase chain reaction method.The designed six pairs of primers can amplify proper fragments and avoid primer dimers.The PCR conditions for the multiplex PCR analysis were optimized based on the primer concentration and annealing temperature,and the Six-Plex PCR detection system for transgenic components in cotton was then established.Results showed that the Six-Plex PCR can successfully detect the transgenic components in genetically modified cotton,from even a small quantity sample as little as a cotton seed using a modified genomic DNA extraction method for cotton seeds in this study. |
| Key words: cotton transgenic detection multiplex PCR |
