期刊
NATURE COMMUNICATIONS
卷 9, 期 -, 页码 -出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/s41467-018-04254-0
关键词
-
资金
- Biotechnology and Biological Sciences Research Council [BB/M005690/1]
- Bill & Melinda Gates Foundation award [RB0447]
- University of Manchester President's Award for Research Excellence
- National Natural Science Foundation of China [31471254]
- National Science Fund for Distinguished Young Scholars [31725002]
- China Postdoctoral Science Foundation [2017M620393]
- Guangdong Provincial Key Laboratory of Genome Read and Write [2017B030301011]
- Autodesk and University of Edinburgh
- Manchester Centre for Synthetic Biology of Fine and Specialty Chemicals (SYNBIOCHEM) [BB/M017702/1]
- Scottish Universities Life Sciences Alliance (SULSA)
- Medical Research Council [J54359]
- Wellcome Trust [201531/Z/16/Z]
- BBSRC [BB/M017702/1, BB/P02114X/1, BB/M005690/1, BB/M025640/1, BB/M005690/2] Funding Source: UKRI
- EPSRC [EP/P017401/1] Funding Source: UKRI
Exogenous pathway optimization and chassis engineering are two crucial methods for heterologous pathway expression. The two methods are normally carried out step-wise and in a trial-and-error manner. Here we report a recombinase-based combinatorial method (termed SCRaMbLE-in) to tackle both challenges simultaneously. SCRaMbLE-in includes an in vitro recombinase toolkit to rapidly prototype and diversify gene expression at the pathway level and an in vivo genome reshuffling system to integrate assembled pathways into the synthetic yeast genome while combinatorially causing massive genome rearrangements in the host chassis. A set of loxP mutant pairs was identified to maximize the efficiency of the in vitro diversification. Exemplar pathways of beta-carotene and violacein were successfully assembled, diversified, and integrated using this SCRaMbLE-in method. High-throughput sequencing was performed on selected engineered strains to reveal the resulting genotypeto-phenotype relationships. The SCRaMbLE-in method proves to be a rapid, efficient, and universal method to fast track the cycle of engineering biology.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据