中文摘要：未知DNA序列的两侧已知序列的分离是转基因生物品系特异性检测的一项重要任务。品系特异性检测方法都不是基于转基因马铃薯AV43-6-G7外源性要素的交接序列。本文通过外源基因5'-RACE成功地获得了该马铃薯外源性片段与重组染色体的侧翼序列。作者利用根据侧翼序列设计的引物建立了转基因马铃薯AV43-6-G7的特异性实时PCR和数字PCR检测方法。研究结果表明，实时PCR和数字PCR方法可用于转基因马铃薯AV43-6-G7的检测。
外文摘要：Background: The isolation of unknown DNA sequences flanked by known sequences is an important task in the event-specific detection of GMOs. None of event-specific detection method was developed based on the junction sequence of an exogenous integrant in the transgenic potato AV43-6-G7. Results: The flanking sequence between the exogenous fragment and recombinant chromosome of this potato was successfully acquired through exogenous gene 5'-RACE. The event-specific primers and Taqman probe were designed to amplify fragments spanning the exogenous DNA and potato genomic DNA. The specific real-time PCR and digital PCR detection methods for AV43-6-G7 potato were established based on primers designed according to the flanking sequences. The detection limit of the qualitative PCR assay was 0.01 % for AV43-6-G7 potato in 100 ng of potato genomic DNA, corresponding to approximately 11.6 copies of the potato haploid genome. The ddPCR assays for Potato AV43-6-G7 achieved a limit of quantification of approximately 58 target copies, with RSD <= 25 %. The aLOQ of this system was approximately 1.2 copies. Conclusions: These results indicated that these event-specific methods would be useful for the identification of potato AV43-6-G7.
外文关键词：Transgenic；Potato AV43-6-G7；Flanking sequence；Event Specific；Real-time PCR；Detection
作者：Gao, Hongwei；Yu, Xiaofan；Deng, Tingting；等
作者单位：Shandong Entry Exit Inspect & Quarantine Bur Peop
期刊名称：BMC BIOTECHNOLOGY
期刊影响因子：2.452
出版年份：2016
出版刊次：10
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