摘要
以转基因小麦B73-6-1为研究对象,通过染色体步行技术,成功分离到B73-6-1上pAHC25质粒外源基因插入位点的3'端旁侧序列,其扩增片段覆盖了转化载体及转基因小麦基因组旁侧序列。同时根据旁侧序列设计引物,建立品系特异性定性PCR检测方法,以典型的转基因作物证明该方法检测B73-6-1具有高特异性。该方法特异性好、灵敏度高,可快速、准确、高效地检测转基因小麦B73-6-1品种。
By Chromosome walking PCR technique,the 3′flanking sequence of the exogenous gene in the pAHC25 plasmid of wheat ＇B73-6-1＇ was successfully cloned.An event specific transgenic detection method for ＇B73-6-1＇ was established by PCR with the specific primers based on the 3′ border.The amplification fragment include part sequence of both the exogenous DNA and wheat genome.This assay was applied to detect typical genetically modified crops,with high specificity for B73-6-1,and the limit of detection was 0.1%.The results showed that the qualitative PCR assay was accurate,rapid and efficient for detection of B73-6-1.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2011年第10期100-105,共6页
China Biotechnology
基金
转基因重大专项(2008ZX08012-001)资助项目
