Institute for Biology I, RWTH Aachen University, Worringer Weg 1, 52056, Aachen, Germany.
Transgenic Res. 2013 Feb;22(1):153-67. doi: 10.1007/s11248-012-9640-0. Epub 2012 Sep 13.
At present, only few methods for the effective assembly of multigene constructs have been described. Here we present an improved version of the MultiRound Gateway technology, which facilitates plant multigene transformation. The system consists of two attL-flanked entry vectors, which contain an attR cassette, and a transformation-competent artificial chromosome based destination vector. By alternate use of the two entry vectors, multiple transgenes can be delivered sequentially into the Gateway-compatible destination vector. Multigene constructs that carried up to seven transgenes corresponding to more than 26 kb were assembled by seven rounds of LR recombination. The constructs were successfully transformed into tobacco plants and were stably inherited for at least two generations. Thus, our system represents a powerful, highly efficient tool for multigene plant transformation and may facilitate genetic engineering of agronomic traits or the assembly of genetic pathways for the production of biofuels, industrial or pharmaceutical compounds in plants.
目前，仅有少数几种有效的多基因构建体组装方法被描述。在这里，我们提出了一种改进的 MultiRound Gateway 技术，该技术可促进植物的多基因转化。该系统由两个侧翼带有 attL 的入口载体组成，它们包含一个 attR 盒和一个基于转化能力的人工染色体的目的载体。通过交替使用这两个入口载体，可以将多个转基因顺次递送到 Gateway 兼容的目的载体中。通过 7 轮 LR 重组，成功地将多达 7 个转基因构建体（对应于超过 26 kb）组装在一起。构建体被成功转化到烟草植物中，并至少稳定遗传了两代。因此，我们的系统代表了一种强大的、高效的多基因植物转化工具，可能有助于农业性状的遗传工程或生物燃料、工业或医药化合物在植物中的遗传途径的组装。
