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| 利用竞争性PCR筛选无标记Xa21转基因水稻 | |
| 引用本文： | 夏志辉,高利芬,罗越华,邓晓建,李仕贵,翟文学.利用竞争性PCR筛选无标记Xa21转基因水稻[J].生物工程学报,2009,25(4):605-610. |
| 作者姓名： | 夏志辉 高利芬 罗越华 邓晓建 李仕贵 翟文学 |
| 作者单位： | 1. 海南大学农学院,海口,570228;中国科学院遗传与发育生物学研究所,北京,100101 2. 中国科学院遗传与发育生物学研究所,北京,100101 3. 海南大学农学院.海口,570228 4. 四川农业大学水稻研究所,成都,611130 |
| 基金项目： | 国家科技部研究项目(Nos.2006AA10Z1C8,2006AA100101,2007BAD81B01);;中国科学院创新项目(Nos.KSCX-YW-N-009-02,KSCX1-YW-03)资助~~ |
| 摘 要： | PCR是一种简单、迅速、灵敏的检测方法,但假阳性与假阴性却影响了它在常规应用中的准确性。本研究利用竞争性PCR解决无标记Xa21转基因水稻PCR检测中的假阳性与假阴性问题。标记基因潮霉素基因(Hygromycin phosphotransferase,hpt)的竞争模板是外加的日本晴hpt转基因植株基因组DNA,抗白叶枯病基因Xa21的竞争模板是待测水稻内源的位于第11染色体上的Xa21同源基因序列。利用这一方法对双右边界T-DNA载体转化产生的转基因T1代植株进行分析,可以有效地减少或排除假阳性或假阴性样品,选出真正的转基因阳性植株。与常规PCR相比竞争性PCR提高了无标记Xa21转基因植株筛选的准确性。对获得的无标记Xa21转基因植株进行白叶枯抗病鉴定与潮霉素抗性鉴定证实了该方法的可靠性。 |
| 关 键 词： | 竞争性PCR 假阳性 假阴性 无标记转基因水稻 Xa21 |
| 收稿时间： | 2008/12/19 0:00:00 |
Application of competitive PCR for screening selectable marker-free Xa21 transgenic rice |
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| Zhihui Xi,Lifen Gao,Yuehua Luo,Xiaojian Deng,Shigui Li and Wenxue Zhai.Application of competitive PCR for screening selectable marker-free Xa21 transgenic rice[J].Chinese Journal of Biotechnology,2009,25(4):605-610. | |
| Authors: | Zhihui Xi Lifen Gao Yuehua Luo Xiaojian Deng Shigui Li and Wenxue Zhai |
| Institution: | College of Agriculture, Hainan University, Haikou 570228, China; Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China;College of Agriculture, Hainan University, Haikou 570228, China;Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China;Rice Research Institute, Sichuan Agricultural University, Chengdu 611130, China;Rice Research Institute, Sichuan Agricultural University, Chengdu 611130, China;College of Agriculture, Hainan University, Haikou 570228, China |
| Abstract: | Polymerase chain reaction (PCR) is a simple,quick and highly sensitive method. However the accuracy of the conventional PCR assay was often affected by false positives and false negatives. In this study,a protocol competitive PCR was used to reduce the false results in screening for selectable marker-free (SMF) Xa21 transgenic rice plants. The competitive template of Xa21 was the endogenous Xa21 homologous sequence located on chromosome 11. The competitive template of the selectable marker gene,hygromycin p... |
| Keywords: | competitive PCR false positive false negative selectable marker-free transgenic rice Xa21 |
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