CAJ | 학술논문

采用基因组步移法和巢式PCR方法研究转基因玉米MON 88017外源基因插入位点旁侧序列特征,获得了外源基因插入位点的左边界旁侧序列504 bp,包括336 bp的插入载体序列和168 bp的玉米基因组序列.根据此旁侧序列,设计MON 88017转化事件特异性引物并进行定性PCR扩增,扩增片段为446 bp.建市了转基因玉米MON88017转化体特异性检测方法.该方法特异性强、灵敏度高(0.1%),为转基因玉米品种MON 88017进出口检测和标识检测提供了技术基础.
채용기인조보이법화소식PCR방법연구전기인옥미MON 88017외원기인삽입위점방측서렬특정,획득료외원기인삽입위점적좌변계방측서렬504 bp,포괄336 bp적삽입재체서렬화168 bp적옥미기인조서렬.근거차방측서렬,설계MON 88017전화사건특이성인물병진행정성PCR확증,확증편단위446 bp.건시료전기인옥미MON88017전화체특이성검측방법.해방법특이성강、령민도고(0.1%),위전기인옥미품충MON 88017진출구검측화표식검측제공료기술기출.
The experiment was conducted to investigate the integration site of transgene in maize MON88017 and to establish event specific methods for qualitative detection of MON88017 based on the left border junction fragment, which was isolated with the amended GenomeWalker and Nested-PCR methods. Sequence alignment between the T-DNA sequence and isolated junction fragments showed a 504 bp junction fragment of MON88017 including 336 bp of T-DNA sequence and 168 bp of MON88017 genome DNA. MON88017 event-specific qualitative PCR method was established with the primers (MON88017-1F/R) targeting the junction regions to produce a 446 bp product. The limit of detection for qualitative PCR assay was 0.1%. The method developed in this work is highly specific, sensitive and suitable for MON88017 sample detection.