Gils Mario, Rubtsova Myroslava, Kempe Katja
Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK) Gatersleben, Gatersleben, Germany.
Methods Mol Biol. 2012;847:123-35. doi: 10.1007/978-1-61779-558-9_11.
The establishment of traits that result from the concerted expression of complementing transgene fragments is a feasible tool for trait control or gene flow control in plants. This chapter describes the methodology for producing herbicide-resistant and pollen-sterile wheat plants by the intein-mediated assembly of inactive precursor protein fragments (protein trans-splicing). We suggest the design of intein-containing vectors for split-transgene expression. We describe transient plant assays that can be used to analyse the functionality of the system and describe the transformation of wheat plants using a split selection marker.We hope that this chapter will be a helpful guideline for researchers who are interested in applying similar split-gene approaches in wheat or other monocotyledonous crops.
由互补转基因片段协同表达产生的性状确立,是植物性状控制或基因流控制的一种可行工具。本章描述了通过内含肽介导的无活性前体蛋白片段组装(蛋白质反式剪接)来生产抗除草剂和花粉不育小麦植株的方法。我们建议设计用于分裂转基因表达的含内含肽载体。我们描述了可用于分析该系统功能的瞬时植物检测,并描述了使用分裂选择标记对小麦植株进行转化的方法。我们希望本章能为有兴趣在小麦或其他单子叶作物中应用类似分裂基因方法的研究人员提供有益的指导。