| 抗虫转CpTI基因植物定性PCR检测技术的建立 |
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| 引用本文： | 朱珠,王永,兰青阔,赵新,余景会,郭永泽,程奕.抗虫转CpTI基因植物定性PCR检测技术的建立[J].河南农业科学,2012,41(5):56-60. |
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| 作者姓名： | 朱珠 王永 兰青阔 赵新 余景会 郭永泽 程奕 |
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| 作者单位： | 天津市农业科学院中心实验室,天津,300381 |
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| 摘 要： | 为建立CpTI基因定性PCR检测方法,通过数据库查询和测序获得CpTI基因序列,设计特异性定性PCR引物,对引物、PCR反应条件和反应体系进行优化,并对检测方法的特异性、灵敏度等进行分析试验。结果表明,CpTI基因特异性定性PCR检测方法选择退火温度为58℃,引物浓度为0.4μmol/L较好,该方法能够特异性检测样品中CpTI基因,检测灵敏度达0.05%(质量比)。该方法符合农业转基因生物产品成分检测标准对特异性、灵敏度、重复性的要求,可为抗虫转CpTI基因植物生物安全管理提供技术支撑。
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| 关 键 词： | 豇豆胰蛋白酶抑制剂 基因特异性检测 定性PCR |
Cowpea Trypsin Inhibitor (CpTI) Gene-specific Qualitative PCR Method for Detection of the Insect Resistance CpTI Gene Plants |
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| ZHU Zhu
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WANG Yong
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LAN Qing-kuo
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ZHAO Xin
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YU Jing-hui
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GUO Yong-ze
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CHENG Yi.Cowpea Trypsin Inhibitor (CpTI) Gene-specific Qualitative PCR Method for Detection of the Insect Resistance CpTI Gene Plants[J].Journal of Henan Agricultural Sciences,2012,41(5):56-60. |
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| Authors: | ZHU Zhu WANG Yong LAN Qing-kuo ZHAO Xin YU Jing-hui GUO Yong-ze CHENG Yi |
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| Institution: | (Central Lab of Tianjin Agriculture Academy Sciences,Tianjin 300381,China) |
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| Abstract: | To develop the qualitative PCR assay for detection cowpea trypsin inhibitor(CpTI) gene,a series of gene-specific qualitative PCR primers were designed according to the sequences of CpTI gene from GenBank and Sanger sequencing.The annealing temperature of primers,PCR reaction system and PCR amplification conditions were optimized,and the specificity and sensitivity of reactions were tested.The results showed that the optimal annealing temperature was 58 ℃ and the optimal primer concentration was 0.4 μmol/L.The CpTI gene could be distinguished specifically among the samples by this method,and the limit of detection sensitivity was 0.05%(w/w),which accorded with the request of the genetically modified organisms(GMO) testing standard for specificity,sensitivity and repeatability.This detection method provided the corresponding technology support to the genetically modified organisms safety management of insect-resistant plant with CpTI gene. |
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| Keywords: | cowpea trypsin inhibitor(CpTI) gene-specific test qualitative PCR |
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