Development of Terminator–Promoter Bifunctional Elements for Application in <i>Saccharomyces cerevisiae</i> Pathway Engineering
The construction of a genetic circuit requires the substitution and redesign of different promoters and terminators. The assembly efficiency of exogenous pathways will also decrease significantly when the number of regulatory elements and genes is increased. We speculated that a novel bifunctional e...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-06-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/12/9870 |
| _version_ | 1797594328949850112 |
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| author | Xiaoxia Ni Zhengyang Liu Jintang Guo Genlin Zhang |
| author_facet | Xiaoxia Ni Zhengyang Liu Jintang Guo Genlin Zhang |
| author_sort | Xiaoxia Ni |
| collection | DOAJ |
| description | The construction of a genetic circuit requires the substitution and redesign of different promoters and terminators. The assembly efficiency of exogenous pathways will also decrease significantly when the number of regulatory elements and genes is increased. We speculated that a novel bifunctional element with promoter and terminator functions could be created via the fusion of a termination signal with a promoter sequence. In this study, the elements from a <i>Saccharomyces cerevisiae</i> promoter and terminator were employed to design a synthetic bifunctional element. The promoter strength of the synthetic element is apparently regulated through a spacer sequence and an upstream activating sequence (UAS) with a ~5-fold increase, and the terminator strength could be finely regulated by the efficiency element, with a ~5-fold increase. Furthermore, the use of a TATA box-like sequence resulted in the adequate execution of both functions of the TATA box and the efficiency element. By regulating the TATA box-like sequence, UAS, and spacer sequence, the strengths of the promoter-like and terminator-like bifunctional elements were optimally fine-tuned with ~8-fold and ~7-fold increases, respectively. The application of bifunctional elements in the lycopene biosynthetic pathway showed an improved pathway assembly efficiency and higher lycopene yield. The designed bifunctional elements effectively simplified pathway construction and can serve as a useful toolbox for yeast synthetic biology. |
| first_indexed | 2024-03-11T02:22:07Z |
| format | Article |
| id | doaj.art-1de1c878ae3443fbbcb36cd6c1d34e24 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T02:22:07Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-1de1c878ae3443fbbcb36cd6c1d34e242023-11-18T10:45:18ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-06-012412987010.3390/ijms24129870Development of Terminator–Promoter Bifunctional Elements for Application in <i>Saccharomyces cerevisiae</i> Pathway EngineeringXiaoxia Ni0Zhengyang Liu1Jintang Guo2Genlin Zhang3Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, Shihezi 832003, ChinaKey Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, Shihezi 832003, ChinaKey Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, Shihezi 832003, ChinaKey Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, Shihezi 832003, ChinaThe construction of a genetic circuit requires the substitution and redesign of different promoters and terminators. The assembly efficiency of exogenous pathways will also decrease significantly when the number of regulatory elements and genes is increased. We speculated that a novel bifunctional element with promoter and terminator functions could be created via the fusion of a termination signal with a promoter sequence. In this study, the elements from a <i>Saccharomyces cerevisiae</i> promoter and terminator were employed to design a synthetic bifunctional element. The promoter strength of the synthetic element is apparently regulated through a spacer sequence and an upstream activating sequence (UAS) with a ~5-fold increase, and the terminator strength could be finely regulated by the efficiency element, with a ~5-fold increase. Furthermore, the use of a TATA box-like sequence resulted in the adequate execution of both functions of the TATA box and the efficiency element. By regulating the TATA box-like sequence, UAS, and spacer sequence, the strengths of the promoter-like and terminator-like bifunctional elements were optimally fine-tuned with ~8-fold and ~7-fold increases, respectively. The application of bifunctional elements in the lycopene biosynthetic pathway showed an improved pathway assembly efficiency and higher lycopene yield. The designed bifunctional elements effectively simplified pathway construction and can serve as a useful toolbox for yeast synthetic biology.https://www.mdpi.com/1422-0067/24/12/9870synthetic regulatory elementpathway engineeringpromoterterminatortranscriptional regulation |
| spellingShingle | Xiaoxia Ni Zhengyang Liu Jintang Guo Genlin Zhang Development of Terminator–Promoter Bifunctional Elements for Application in <i>Saccharomyces cerevisiae</i> Pathway Engineering International Journal of Molecular Sciences synthetic regulatory element pathway engineering promoter terminator transcriptional regulation |
| title | Development of Terminator–Promoter Bifunctional Elements for Application in <i>Saccharomyces cerevisiae</i> Pathway Engineering |
| title_full | Development of Terminator–Promoter Bifunctional Elements for Application in <i>Saccharomyces cerevisiae</i> Pathway Engineering |
| title_fullStr | Development of Terminator–Promoter Bifunctional Elements for Application in <i>Saccharomyces cerevisiae</i> Pathway Engineering |
| title_full_unstemmed | Development of Terminator–Promoter Bifunctional Elements for Application in <i>Saccharomyces cerevisiae</i> Pathway Engineering |
| title_short | Development of Terminator–Promoter Bifunctional Elements for Application in <i>Saccharomyces cerevisiae</i> Pathway Engineering |
| title_sort | development of terminator promoter bifunctional elements for application in i saccharomyces cerevisiae i pathway engineering |
| topic | synthetic regulatory element pathway engineering promoter terminator transcriptional regulation |
| url | https://www.mdpi.com/1422-0067/24/12/9870 |
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