DNA Assembly via Reiterative Recombination

描述(由申请人提供)：代谢工程已成为生产具有重要治疗作用的天然产物及其类似物的一种很有前途的策略。然而，生物合成基因途径的构建和操纵构成了工程细胞生产复杂天然产物及其衍生物的关键瓶颈，需要新一代变革性技术来克服这一障碍。为此，我们开发了叠加性重组作为一种策略，用于在体内有效地组装多基因DNA结构。在这里，我们建议开发我们的代谢工程应用技术，在优化酵母中紫杉醇中间体的萜类产量的背景下挑战它。如果成功，这项研究将 使天然产物生物合成途径库的构建成为一种常规的、负担得起的、高效的过程，非专家可以在没有任何专门设备的情况下进行。

DESCRIPTION (provided by applicant): Metabolic engineering has emerged as a promising strategy for the production of therapeutically important natural products and their analogs. However, the construction and manipulation of pathways of biosynthetic genes poses a key bottleneck to efforts to engineer cells for the production of complex natural products and their derivatives, and a new generation of transformative technologies will be needed to overcome this barrier. Towards this end, we have developed Reiterative Recombination as a strategy for the efficient in vivo assembly of multi-gene DNA constructs. Here we propose to develop our technology for metabolic engineering applications by challenging it in the context of optimizing terpenoid yields, specifically for taxol intermediates, in yeast. If successful, this research will make the construction of libraries of natural product biosynthetic pathways a routine, affordable, and highly efficient process that can be carried out by the non-expert without any specialized equipment.