CP4-EPSPS转基因棉花植株鉴定方法比较分析

发布时间：2018-06-04 00:31

本文选题：转基因棉花 + CP-EPSPS　；参考：《生物技术通报》2017年04期

【摘要】：CP4-EPSPS是从土壤农杆菌CP4菌株中分离得到的抗除草剂草甘膦基因,是转基因植物研究中最为常用的一种筛选标记基因。但是在CP4-EPSPS转基因植物后期筛选过程中仅凭简单的草甘膦抗性筛选并不能得到完全准确的鉴定结果,常会出现假阳性或漏选的情况。本研究以抗草甘膦转CP4-EPSPS基因棉花为材料,对PCR扩增、ELISA、CP4-EPSPS蛋白检测试纸条及草甘膦抗性四种转基因阳性植株筛选方法进行比较,从筛选结果的准确性、时效性、易用性及实惠性等不同方面综合分析了各种筛选方法的优缺点,结果表明ELISA、试纸条检测方法的准确性显著高于PCR检测与0.3%浓度草甘膦抗性检测,而试纸条检测操作更加便捷,因此建议将草甘膦抗性与试纸条检测方法结合起来使用。本研究结果为以CP4-EPSPS基因为标记筛选的转基因植物的室内及大田筛选提供实验依据。
[Abstract]:CP4-EPSPS is a herbicide resistant glyphosate gene isolated from soil Agrobacterium tumefaciens CP4. It is one of the most commonly used screening marker genes in transgenic plant studies. However, in the late screening process of CP4-EPSPS transgenic plants, only simple glyphosate resistance can not be obtained by simple glyphosate resistance. False positive or leaky selection. In this study, CP4-EPSPS transgenic cotton was used as the material. The screening methods of PCR amplification, ELISA, CP4-EPSPS protein test paper and glyphosate resistance four transgenic positive plants were compared. The results were analyzed from different aspects, such as accuracy, timeliness, ease of use and benefits. The advantages and disadvantages of the screening method show that the accuracy of the test paper detection method is significantly higher than that of PCR detection and 0.3% concentration of glyphosate resistance detection, and the test strip detection operation is more convenient. Therefore, it is suggested to combine the glyphosate resistance with the test paper detection method. The results of this study are selected by the CP4-EPSPS gene as a marker. Laboratory and field screening of transgenic plants provides experimental evidence.
【作者单位】：中国农业科学院生物技术研究所;
【基金】：国家转基因生物新品种培育科技重大专项资助项目(2016ZX08005-004)
【分类号】：S562

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