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

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-synthe...

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Autores principales:	Wen Wan, Min Lu, Dongmei Wang, Xiaolian Gao, Jiong Hong
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2141704e9cb34e5daad9427306b9b4d5
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spelling	oai:doaj.org-article:2141704e9cb34e5daad9427306b9b4d52021-12-02T12:32:15ZHigh-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment10.1038/s41598-017-06428-02045-2322https://doaj.org/article/2141704e9cb34e5daad9427306b9b4d52017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-06428-0https://doaj.org/toc/2045-2322Abstract 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.Wen WanMin LuDongmei WangXiaolian GaoJiong HongNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Wen Wan Min Lu Dongmei Wang Xiaolian Gao Jiong Hong High-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment
description	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.
format	article
author	Wen Wan Min Lu Dongmei Wang Xiaolian Gao Jiong Hong
author_facet	Wen Wan Min Lu Dongmei Wang Xiaolian Gao Jiong Hong
author_sort	Wen Wan
title	High-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment
title_short	High-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment
title_full	High-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment
title_fullStr	High-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment
title_full_unstemmed	High-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment
title_sort	high-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/2141704e9cb34e5daad9427306b9b4d5
work_keys_str_mv	AT wenwan highfidelitydenovosynthesisofpathwaysusingmicrochipsynthesizedoligonucleotidesandgeneralmolecularbiologyequipment AT minlu highfidelitydenovosynthesisofpathwaysusingmicrochipsynthesizedoligonucleotidesandgeneralmolecularbiologyequipment AT dongmeiwang highfidelitydenovosynthesisofpathwaysusingmicrochipsynthesizedoligonucleotidesandgeneralmolecularbiologyequipment AT xiaoliangao highfidelitydenovosynthesisofpathwaysusingmicrochipsynthesizedoligonucleotidesandgeneralmolecularbiologyequipment AT jionghong highfidelitydenovosynthesisofpathwaysusingmicrochipsynthesizedoligonucleotidesandgeneralmolecularbiologyequipment
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High-fidelity de novo synthesis of pathways using microchip-synthesized oligonucleotides and general molecular biology equipment

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