| Inverse PCR法快速测定转基因马铃薯中T-DNA的拷贝数 |
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| 引用本文： | 南相日.Inverse PCR法快速测定转基因马铃薯中T-DNA的拷贝数[J].中国马铃薯,2006,20(2):78-80. |
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| 作者姓名： | 南相日 |
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| 作者单位： | 黑龙江省农业科学院生物技术研究中心,黑龙江,哈尔滨,150086 |
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| 摘 要： | 利用Inverse PCR技术分析原生质体途径导入PLRV-CP基因的马铃薯转基因后代T-DNA拷贝数,结果与Southern Blot相同。经RT-PCR分析,低拷贝数的转基因后代显阳性反应,多拷贝数的显阴性反应,说明T-DNA多拷贝数影响外源基因的表达。Inverse PCR技术能为早期快速准确预测转基因植株中外源基因的T-DNA拷贝数,排除多拷贝重复串联序列诱导的外源基因沉默植株,为获得稳定表达的转基因株系提供有效方法。
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| 关 键 词： | Inverse PCR 拷贝数 基因沉默 |
| 文章编号： | 1672-3635(2006)02-078-03 |
| 修稿时间： | 2006年1月23日 |
A Quick Method to Estimate T-DNA Copy Number in Potato Transgenic Plants Using Inverse PCR |
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| Nan Xiangri.A Quick Method to Estimate T-DNA Copy Number in Potato Transgenic Plants Using Inverse PCR[J].Chinese Potato,2006,20(2):78-80. |
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| Authors: | Nan Xiangri |
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| Abstract: | Inverse PCR technique was used to analyze T-DNA copy number in PLRV-CP gene transformed potato offsprings obtained by protoplasts mediated way.The results were identical to Southern Blot.Through RT-PCR analysis,low copy number transgenic plants showed positive reaction and multi-copy number showed negative reaction.It was suggested that the expression of foreign gene was influenced by the T-DNA copy number.Inverse PCR technique provides a quick and efficient way to estimate T-DNA copy number in potato transgenic plants at early stage and eliminate repeat induced gene silencing plants caused by multi-copy T-DNA number,providing a valid method to obtain a stabile expression transgenic plant. |
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| Keywords: | Inverse PCR |
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