转基因香石竹Moonlite品系特异性定性PCR检测方法的建立

Establishment of Event-specific Qualitative PCR for Detecting Transgenic Carnation 'Moonlite'

以转基因香石竹品种Moonlite(月之伊人)为研究对象,通过TAIL-PCR方法,获得外源基因插入位点的左侧旁邻序列。根据旁邻序列设计引物,建立品系特异性定性PCR方法,其扩增片段覆盖了转化载体及香石竹基因组临界序列。本方法特异性好、灵敏度高,可快速、准确、高效地检测转基因香石竹Moonlite品种。

An event-specific transgenic detection method for ’Moonlite’ was established.By thermal asymmetric interlaced PCR（TAIL-PCR） technique,the flanking sequences of the exogenous integration in the genome of ’Moonlite’ were cloned and sequenced with the specific nested primers based on the left border.According to the left flanking sequence,the event-specific primers were designed to amplify the fragments,which could span the exogenous DNA and carnation genome and the event-specific qualitative PCR detection method for transgenic carnation ’Moonlite’ was established.The results showed that the qualitative PCR assay was accurate,rapid and efficient for detection of ’Moonlite’.