转基因水稻BarKasalath-01事件特异性检测

发布时间：2018-05-04 23:00

本文选题：转基因水稻 + 旁侧序列　；参考：《分子植物育种》2017年11期

【摘要】：利用hi TAIL-PCR方法研究了转基因水稻Bar Kasalath-01中外源基因在基因组上插入位点的序列特征,获得了外源基因T-DNA右边界旁侧序列511 bp,与水稻基因组数据库比对发现,外源基因插入位点位于水稻基因组第8号染色体的第3 326 720处。根据整合位点水稻基因组序列和外源基因T-DNA左边界序列设计引物,扩增得到了左旁侧序列783 bp。基于左右旁侧序列,建立了转基因水稻Bar Kasalath-01的事件特异性定性PCR检测方法,扩增片段分别为783 bp和411 bp。该方法特异性强、灵敏度高,能够在Bar Kasalath-01基因组DNA含量为0.1%的模板中检测出转基因成份。依据旁侧序列,还建立了快速鉴定转基因后代植株基因型的3引物PCR检测方法。这些方法的建立,实现了对转基因水稻Bar Kasalath-01转化事件的特异性检测。
[Abstract]:Hi TAIL-PCR method was used to study the sequence characteristics of the insertion site of Bar Kasalath-01 gene in transgenic rice genome. The sequence of 511 BP on the right side of the T-DNA was obtained, and the sequence was compared with the rice genome database. The insertion site of exogenous gene was located at 3326 720 on chromosome 8 of rice genome. According to the sequence of rice genome and the left bound sequence of exogenous gene T-DNA, primers were designed, and the left flanking sequence of 783 BP was obtained. Based on the left and right flanking sequences, an event-specific qualitative PCR detection method for Bar Kasalath-01 of transgenic rice was established. The amplified fragments were 783bp and 411bprespectively. The method is specific and sensitive, and can be used to detect transgenic components in the template containing 0.1% genomic DNA of Bar Kasalath-01. According to the flanking sequence, a 3 primer PCR method was established for rapid genotyping of transgenic progenies. These methods were used to detect Bar Kasalath-01 transformation events in transgenic rice.
【作者单位】：中国科学院亚热带农业生态研究所亚热带农业生态过程重点实验室;中国科学院大学;湖南省蚕桑科学研究所;
【基金】：转基因生物新品种培育科技重大专项(2016ZX08001-003)资助
【分类号】：S511

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