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。
Inverse PCR technique was used to analyze potato offsprings obtained by protoplasts mediated way. The T-DNA copy number in PLRV-CP gene transformed 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.
出处
《中国马铃薯》
2006年第2期78-80,共3页
Chinese Potato Journal
基金
"十五"黑龙江省攻关项目(GA010B101-08)
