| 地高辛随机引物法标记探针的Southern杂交技术优化 |
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| 引用本文： | 周长发,张锐,张晓,罗淑萍,郭三堆. 地高辛随机引物法标记探针的Southern杂交技术优化[J]. 中国农业科技导报, 2009, 11(4): 123-128 |
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| 作者姓名： | 周长发 张锐 张晓 罗淑萍 郭三堆 |
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| 作者单位： | 1.中国农业科学院生物技术研究所, 农作物基因资源与基因改良国家重大科学工程, 北京 100081; 2.新疆农业大学农学院, 乌鲁木齐 830052 |
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| 基金项目： | 国家转基因生物新品种培育科技重大专项 |
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| 摘 要： | 为检测转基因抗虫棉中的Bt Cry1A基因的表达，采用地高辛随机引物法标记探针、化学发光检测的Southern杂交技术，通过纯化探针、使用最适探针浓度、优化真空转印、杂交及免疫检测方法等一系列优化过程，最终得到背景低、信号强的Southern杂交结果。
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| 关 键 词： | 地高辛 化学发光法检测 转基因作物, |
| 收稿时间： | 2009-04-29 |
| 修稿时间： | 2009-06-30 |
Technological Improvement of Southern Blot Using Digoxigenin-labeled Probes with Random-primed Method |
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| ZHOU Zhang-fa,ZHANG Rui,ZHANG Xiao,LUO Shu-ping,GUO San-dui. Technological Improvement of Southern Blot Using Digoxigenin-labeled Probes with Random-primed Method[J]. Journal of Agricultural Science and Technology, 2009, 11(4): 123-128 |
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| Authors: | ZHOU Zhang-fa ZHANG Rui ZHANG Xiao LUO Shu-ping GUO San-dui |
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| Affiliation: | 1.Biotechnology Research Institute, National Key Facility of Crop Gene Resources and Genetic Improvement, ;Chinese Academy of Agricultural Sciences, Beijing 100081|2.Xinjiang Agricultural University, College of Agriculture, Urumqi 830052,China |
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| Abstract: | Southern blot using digoxigenin(DIG)-labeled probes with random-primed method and chemiluminescent detection was used to detect Bt Cry1A gene of transgenic insect-resistant cotton. A serial of procedure was modified by purifying the DIG-labeled probes, using optimum probes concentration, improving the process of vacuum transferring, hybridization and immunoassay. Finally, we achieved good Southern blot result with low background and high sensitivity. |
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| Keywords: | digoxigenin chemiluminescent detection transgenic plant |
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