One-step construction of a food-grade expression system based on the URA3 gene in Kluyveromyces lactis
Copyright © 2021. Published by Elsevier Inc..
Proteins from food-grade expression systems can be used in food products and medical applications. Herein, we describe a one-step method of constructing an expression vector in Kluyveromyces lactis by combining a URA3-deficient strain and a plasmid vector with no drug-resistant selection. Adjacent DNA elements of the vector were assembled in a targeted manner through a reaction with a special recombinase to form a plasmid vector using a one-step reaction. The unnecessary fragments containing the pUC origin and the ampicillin resistance gene were removed, and the vector was isolated and purified before transformation. A single transformation of the vector can produce a URA3-deficient strain. PCR assay, sequencing, and western blot analysis all indicated that the method of vector construction and target protein expression (mCherry and human serum albumin) were successful. This method may potentially be applied to any species containing the URA3 gene; this system has the potential to become a safe and powerful tool for promoting protein expression in food-safe species.
Medienart: |
E-Artikel |
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Erscheinungsjahr: |
2021 |
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Erschienen: |
2021 |
Enthalten in: |
Zur Gesamtaufnahme - volume:116 |
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Enthalten in: |
Plasmid - 116(2021) vom: 07. Juli, Seite 102577 |
Sprache: |
Englisch |
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Beteiligte Personen: |
Liang, Zhicheng [VerfasserIn] |
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Links: |
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Themen: |
Food-grade expression |
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Anmerkungen: |
Date Completed 28.10.2021 Date Revised 28.10.2021 published: Print-Electronic Citation Status MEDLINE |
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doi: |
10.1016/j.plasmid.2021.102577 |
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funding: |
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Förderinstitution / Projekttitel: |
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PPN (Katalog-ID): |
NLM32606852X |
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520 | |a Copyright © 2021. Published by Elsevier Inc. | ||
520 | |a Proteins from food-grade expression systems can be used in food products and medical applications. Herein, we describe a one-step method of constructing an expression vector in Kluyveromyces lactis by combining a URA3-deficient strain and a plasmid vector with no drug-resistant selection. Adjacent DNA elements of the vector were assembled in a targeted manner through a reaction with a special recombinase to form a plasmid vector using a one-step reaction. The unnecessary fragments containing the pUC origin and the ampicillin resistance gene were removed, and the vector was isolated and purified before transformation. A single transformation of the vector can produce a URA3-deficient strain. PCR assay, sequencing, and western blot analysis all indicated that the method of vector construction and target protein expression (mCherry and human serum albumin) were successful. This method may potentially be applied to any species containing the URA3 gene; this system has the potential to become a safe and powerful tool for promoting protein expression in food-safe species | ||
650 | 4 | |a Journal Article | |
650 | 4 | |a Research Support, Non-U.S. Gov't | |
650 | 4 | |a Food-grade expression | |
650 | 4 | |a Human serum albumin | |
650 | 4 | |a Kluyveromyces lactis | |
650 | 4 | |a URA3 gene | |
650 | 4 | |a mCherry | |
700 | 1 | |a Deng, Mulan |e verfasserin |4 aut | |
700 | 1 | |a Zhang, Zhi |e verfasserin |4 aut | |
700 | 1 | |a Li, Meirong |e verfasserin |4 aut | |
700 | 1 | |a Zhou, SuJin |e verfasserin |4 aut | |
700 | 1 | |a Zhao, ZhengGang |e verfasserin |4 aut | |
700 | 1 | |a Mu, YunPing |e verfasserin |4 aut | |
700 | 1 | |a Wang, LiNa |e verfasserin |4 aut | |
700 | 1 | |a Ning, Chengyun |e verfasserin |4 aut | |
700 | 1 | |a Zhao, Allan Zijian |e verfasserin |4 aut | |
700 | 1 | |a Li, Fanghong |e verfasserin |4 aut | |
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