摘要
以EH92-527-1转基因马铃薯为试材,应用多重PCR技术同时扩增马铃薯的内源基因(UGPase)和外源基因(NOS终止子、NPTII结构基因和EH92-527-1品系特异基因),将扩增产物在DHPLC非变性条件下分离,分析几个基因扩增的结果;将模板进行稀释,确定了方法的检测灵敏度,并与凝胶电泳结果相比较,建立了马铃薯转基因成分筛选检测及品系鉴定的多重PCR-变性高效液相色谱(DHPLC)方法。试验结果表明,该方法能够同时筛选检测转基因马铃薯的四个内、外源基因,且与凝胶成像相比,有更好的检测灵敏度,可达到1 ng/μL。本文首次建立的马铃薯多重PCR-DHPLC检测方法,能够高通量快速准确的检测马铃薯中的转基因成分及对品系进行鉴定。
UGPase gene,EH92-527-1 differential line gene,NOS terminator and NPTII gene in the line EH92-527-1 GMO potato were amplified by multiplex PCR.And the products were separated by DHPLC（Denatured High Performance Liquid Chromatography） undenaturedly.Various grads of samples were used for the sensitivity testing.These results were compared with gel electrophoresis.The multiplex PCR-DHPLC separation method was proposed for the detection of transgenic components and differential line gene in potato.The results showed that the method could be used to detect the four genes in GMO potato.And the limit of detection was 1 ng / μL which was better than that of gel electrophoresis.This method was first-done,rapid,accurate and high throughout.And it could be used for detecting and identifying GMO potato.
出处
《中国马铃薯》
2011年第3期129-134,共6页
Chinese Potato Journal
基金
国家质检总局2011年项目(2011IK200)
黑龙江省科技厅青年基金项目(QC2010022)
