Julius Kuehn Institute, Federal Research Centre for Cultivated Plants (JKI), Institute for Biosafety of Genetically Modified Plants, Erwin-Baur-Str 27, 06484 Quedlinburg, Germany.
Transgenic Res. 2012 Jun;21(3):645-54. doi: 10.1007/s11248-011-9558-y. Epub 2011 Sep 27.
Trait genes are usually introduced into the plant genome together with a marker gene. The last one becomes unnecessary after transgene selection and characterization. One of the strategies to produce transgenic plants free from the selectable marker is based on site-specific recombination. The present study employed the transient Cre-lox system to remove the nptII marker gene from potato. Transient marker gene excision involves introduction of Cre protein in lox-target plants by PVX virus vector followed by plant regeneration. Using optimized experimental conditions, such as particle bombardment infection method and application of P19 silencing suppressor protein, 20-27% of regenerated plants were identified by PCR analysis as marker-free. Based on our comparison of the recombination frequencies observed in this study to the efficiency of other methods to avoid or eliminate marker genes in potato, we suggest that PVX-Cre mediated site-specific excisional recombination is a useful tool to generate potato plants without superfluous transgenic sequences.
性状基因通常与标记基因一起被导入植物基因组。在转基因选择和鉴定后,后者就变得不再需要了。生产不含选择标记的转基因植物的策略之一是基于位点特异性重组。本研究利用瞬时 Cre-lox 系统从马铃薯中去除 nptII 标记基因。瞬时标记基因的切除涉及通过 PVX 病毒载体将 Cre 蛋白引入 lox 靶植物,然后进行植物再生。使用优化的实验条件,例如粒子轰击感染方法和 P19 沉默抑制蛋白的应用,通过 PCR 分析鉴定出 20-27%的再生植物为无标记基因。基于我们将本研究中观察到的重组频率与其他避免或消除马铃薯中标记基因的方法的效率进行比较,我们建议 PVX-Cre 介导的位点特异性切除重组是一种生成无多余转基因序列的马铃薯植物的有用工具。