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

作者：Wan, Wen; Lu, Min; Wang, Dongmei; Gao, Xiaolian; Hong, Jiong^*

来源：Scientific Reports, 2017, 7(1): 6119.

DOI：10.1038/s41598-017-06428-0

• MISMATCH CLEAVAGE
• ERROR-CORRECTION
• DNA-SYNTHESIS
• PCR
• OPTIMIZATION
• RETRIEVAL
• DNAWORKS
• LYCOPENE
• REMOVAL

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.

• 出版日期2017-7-21
• 单位中国科学技术大学; 江苏师范大学