Publications
Scientific reportsJul 2017 |
7
(
1
),
6119
DOI:
10.1038/s41598-017-06428-0

High-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment

Wan, Wen; Lu, Min; Wang, Dongmei; Gao, Xiaolian; Hong, Jiong
Product Used
Oligo Pools
Abstract
Engineering and evaluation of synthetic routes for generating valuable compounds require accurate and cost-effective de novo synthesis of genetic pathways. Here, we present an economical and streamlined de novo DNA synthesis approach for engineering a synthetic pathway with microchip-synthesized oligonucleotides (oligo). The process integrates entire oligo pool amplification, error-removal, and assembly of long DNA molecules. We utilized this method to construct a functional lycopene biosynthetic pathway (11.9 kb encoding 10 genes) in Escherichia coli using a highly error-prone microchip-synthesized oligo pool (479 oligos) without pre-purification, and the error-frequency was reduced from 14.25/kb to 0.53/kb. This low-equipment-dependent and cost-effective method can be widely applied for rapid synthesis of biosynthetic pathways in general molecular biology laboratories.
Product Used
Oligo Pools

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