Department of Plant Pathology, Ohio State University, Columbus OH 43210, USA.
J Integr Plant Biol. 2009 Nov;51(11):982-92. doi: 10.1111/j.1744-7909.2009.00870.x.
Transposons are effective mutagens alternative to T-DNA for the generation of insertional mutants in many plant species including those whose transformation is inefficient. The current strategies of transposon tagging are usually slow and labor-intensive and yield low frequency of tagged lines. We have constructed a series of transposon tagging vectors based on three approaches: (i) AcTPase controlled by glucocorticoid binding domain/VP16 acidic activation domain/Gal4 DNA-binding domain (GVG) chemical-inducible expression system; (ii) deletion of AcTPase via Cre-lox site-specific recombination that was initially triggered by Ds excision; and (iii) suppression of early transposition events in transformed rice callus through a dual-functional hygromycin resistance gene in a novel Ds element (HPT-Ds). We tested these vectors in transgenic rice and characterized the transposition events. Our results showed that these vectors are useful resources for functional genomics of rice and other crop plants. The vectors are freely available for the community.
转座子是有效的诱变剂，可替代 T-DNA 在许多植物物种中产生插入突变体，包括那些转化效率低的物种。目前的转座子标签策略通常较慢且劳动强度大，并且标记线的频率较低。我们构建了一系列基于三种方法的转座子标签载体：（i）受糖皮质激素结合域/VP16 酸性激活域/ Gal4 DNA 结合域（GVG）化学诱导表达系统控制的 AcTPase；（ii）通过 Cre-lox 位点特异性重组删除 AcTPase，最初由 Ds 切除引发；（iii）通过新型 Ds 元件（HPT-Ds）中的双功能潮霉素抗性基因抑制转化水稻愈伤组织中的早期转座事件。我们在转基因水稻中测试了这些载体，并对转座事件进行了表征。我们的结果表明，这些载体是水稻和其他作物功能基因组学的有用资源。这些载体可供社区免费使用。
