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| 一种快速鉴定转基因植物纯合体的新方法 | |
| 引用本文： | 杜春芳,李朋波,李润植. 一种快速鉴定转基因植物纯合体的新方法[J]. 生物技术通讯, 2004, 15(6): 585-587 |
| 作者姓名： | 杜春芳 李朋波 李润植 |
| 作者单位： | 1. 山西省农业科学院,棉花研究所,运城,044000;山西农业大学,农业生物工程研究中心,太谷,030801 2. 山西省农业科学院,棉花研究所,运城,044000 3. 山西农业大学,农业生物工程研究中心,太谷,030801 |
| 基金项目： | 教育部科技重点项目和归国留学人员科研基金项目 |
| 摘 要： | 植物转化中鉴定转基因植物的整合性是一个很重要的步骤，常规方法是对独立分离的转基因T1代植株产生的T2代进行转基因分离比率研究，以检测T1代的转基因整合状态，不仅费时费力，而且浪费了T1代资源。本介绍一种应用双重定量实时PCR技术鉴定转基因植物纯合子的新方法：以T1代植物DNA为模板，根据转基因后代的Ct表型值鉴定其转基因整合状态，Ct值接近2的为转基因纯合型，Ct值接近1的为转基因杂合型。用这种方法，可以同时对数十个T1代转基因幼苗的整合状态进行快速鉴定，准确率为100％。 |
| 关 键 词： | Ct值 快速鉴定 实时PCR 杂合 转基因后代 纯合子 状态 转基因植物 植株 幼苗 |
| 文章编号： | 1009-0002(2004)06-0585-03 |
| 修稿时间： | 2004-06-01 |
A new method for the rapid identification of homozygous transgenic plants |
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| DU Chun-fang,,LI Peng-bo,LI Run-zhi . Cotton Research Institute,Shanxi Academy of Agricultural Sciences,Yuncheng , . Center for Agricultural Biotechnology,Shanxi Agricultural University,Taigu ,China. A new method for the rapid identification of homozygous transgenic plants[J]. Letters in Biotechnology, 2004, 15(6): 585-587 | |
| Authors: | DU Chun-fang LI Peng-bo LI Run-zhi . Cotton Research Institute Shanxi Academy of Agricultural Sciences Yuncheng . Center for Agricultural Biotechnology Shanxi Agricultural University Taigu China |
| Affiliation: | DU Chun-fang1,2,LI Peng-bo1,LI Run-zhi2 1. Cotton Research Institute,Shanxi Academy of Agricultural Sciences,Yuncheng 044000, 2. Center for Agricultural Biotechnology,Shanxi Agricultural University,Taigu 030801,China |
| Abstract: | Identification of zygosity of transgenic plants is an important step for crop transformation. It is a time-consuming and laborious lab work to identify the zygosity of transgenic plants by conventional genetic analysis based on the segregation ratio of transgene among T2 population derived from each independent T1 transgenic plant. This common procedure is often accompanied by the waste of the T1 generation. In this paper, a new method to identify homozygous transgenic plants was described, which employed duplex quantitative real-time PCR. The zygosity of transgenic plants was determined by Ct values of duplex quantitative real-time PCR using T1 plant DNA as templates. Plants with a Ct score of 2 are homozygous for the transgene whereas those scored 1 are heterozygous transgenic plants. The zygosity of several dozen of T1 seedlings was rapidly identified with 100% fidelity using this simple assay. |
| Keywords: | real-time PCR Ct values transgenic plants zygosityy |
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