DNA Assembly via Reiterative Recombination

通过重复重组进行 DNA 组装

基本信息

批准号：
8417300
负责人：
VIRGINIA W CORNISH
金额：
$ 6.76万
依托单位：
COLUMBIA UNIV NEW YORK MORNINGSIDE
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-02-01 至 2016-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8417300
关键词：
Adoption Affect Anabolism Biological Factors Carbon Cell Culture Techniques Cells Chromosomes Codon Nucleotides Commit Communities Complementary DNA Complex Cytochrome P450 DNA Development Double Strand Break Repair Engineering Enzymes Equipment Generations Genes Genetic Genetic Recombination Goals In Vitro Individual Laboratories Length Libraries Metabolic Metabolic Pathway Methods Modeling Molecular Biology Techniques Organism Paclitaxel Pathway interactions Pigments Plants Process Production Recombinants Reporter Genes Research Resources Skeleton Solutions Technology Terpenes Terpenoid Biosynthesis Pathway Testing Therapeutic Time Transplantation Work Yeasts analog base cellular engineering combinatorial design directed evolution empowered feeding homologous recombination improved in vivo isopentenyl pyrophosphate lycopene microbial host microorganism overexpression promoter synthetic biology taxadiene

项目摘要

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.

描述（由申请人提供）：代谢工程已成为生产治疗上重要的天然产物及其类似物的有前景的策略。然而，生物合成基因途径的构建和操作对改造细胞以生产复杂天然产物及其衍生物的努力构成了关键瓶颈，需要新一代的转化技术来克服这一障碍。为此，我们开发了重复重组作为多基因 DNA 构建体体内高效组装的策略。在这里，我们建议通过在酵母中优化萜类化合物产量（特别是紫杉醇中间体）的背景下挑战它来开发我们的代谢工程应用技术。如果成功的话，这项研究将使天然产物生物合成途径文库的构建成为一种常规、负担得起且高效的过程，可由非专家无需任何专门设备即可进行。