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Functional analysis of a viral promoter from a strawberry vein banding virus isolate from China

BACKGROUND: Promoters are important factors affecting gene expression in cells. The driven activities of viral promoters were generally assessed to screen available promoters for transgenic and research and biotech industries. In this study, we cloned a full-length promoter from a Chinese isolate of...

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Autores principales: Jiang, Lei, Chen, Jing, Yang, You-zhi, Li, Rui, Li, Shuang, Wang, Zhan-qi, Jiang, Tong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8974135/
https://www.ncbi.nlm.nih.gov/pubmed/35361243
http://dx.doi.org/10.1186/s12985-022-01778-2
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author Jiang, Lei
Chen, Jing
Yang, You-zhi
Li, Rui
Li, Shuang
Wang, Zhan-qi
Jiang, Tong
author_facet Jiang, Lei
Chen, Jing
Yang, You-zhi
Li, Rui
Li, Shuang
Wang, Zhan-qi
Jiang, Tong
author_sort Jiang, Lei
collection PubMed
description BACKGROUND: Promoters are important factors affecting gene expression in cells. The driven activities of viral promoters were generally assessed to screen available promoters for transgenic and research and biotech industries. In this study, we cloned a full-length promoter from a Chinese isolate of strawberry vein banding virus (SVBV) and produced several deletion mutants for evaluation of applications in production of reporter proteins in stable transgenic plants. METHODS: The full-length promoter of SVBV (SP1) and its three deletion mutants (SP2, SP3, and SP4) were amplified using polymerase chain reaction. The effects of SVBV SP1, SP2, SP3, and SP4 on gene expression were evaluated using β-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes. RESULTS: Transient expression assays showed that the SVBV SP1 promoter and its three deletion mutants all expressed the reporter genes, albeit at very different levels. Interestingly, transcriptional activity driven by the SP1 promoter was much higher than that of the cauliflower mosaic virus (CaMV) 35S promoter. After stable transformation of the GUS gene into Nicotiana tabacum plants, SVBV SP1-driven transgene expression was approximately 2.6-fold higher than CaMV 35S promoter-driven transgene expression. In addition, GUS gene expression levels were enhanced by co-inoculation of the plants with the SP1 promoter-driven vector carrying the GUS gene and the vector expressing SVBV open reading frame (ORF) V or ORF VI. CONCLUSIONS: The SVBV SP1 promoter from the Chinese isolate evaluated in this study could successfully drive transient and stable expression in plants, it was a stronger promoter than the CaMV 35S and FLt-US promoters and may be more useful for the production of stable transgenic plants. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12985-022-01778-2.
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spelling pubmed-89741352022-04-02 Functional analysis of a viral promoter from a strawberry vein banding virus isolate from China Jiang, Lei Chen, Jing Yang, You-zhi Li, Rui Li, Shuang Wang, Zhan-qi Jiang, Tong Virol J Research BACKGROUND: Promoters are important factors affecting gene expression in cells. The driven activities of viral promoters were generally assessed to screen available promoters for transgenic and research and biotech industries. In this study, we cloned a full-length promoter from a Chinese isolate of strawberry vein banding virus (SVBV) and produced several deletion mutants for evaluation of applications in production of reporter proteins in stable transgenic plants. METHODS: The full-length promoter of SVBV (SP1) and its three deletion mutants (SP2, SP3, and SP4) were amplified using polymerase chain reaction. The effects of SVBV SP1, SP2, SP3, and SP4 on gene expression were evaluated using β-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes. RESULTS: Transient expression assays showed that the SVBV SP1 promoter and its three deletion mutants all expressed the reporter genes, albeit at very different levels. Interestingly, transcriptional activity driven by the SP1 promoter was much higher than that of the cauliflower mosaic virus (CaMV) 35S promoter. After stable transformation of the GUS gene into Nicotiana tabacum plants, SVBV SP1-driven transgene expression was approximately 2.6-fold higher than CaMV 35S promoter-driven transgene expression. In addition, GUS gene expression levels were enhanced by co-inoculation of the plants with the SP1 promoter-driven vector carrying the GUS gene and the vector expressing SVBV open reading frame (ORF) V or ORF VI. CONCLUSIONS: The SVBV SP1 promoter from the Chinese isolate evaluated in this study could successfully drive transient and stable expression in plants, it was a stronger promoter than the CaMV 35S and FLt-US promoters and may be more useful for the production of stable transgenic plants. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12985-022-01778-2. BioMed Central 2022-03-31 /pmc/articles/PMC8974135/ /pubmed/35361243 http://dx.doi.org/10.1186/s12985-022-01778-2 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Jiang, Lei
Chen, Jing
Yang, You-zhi
Li, Rui
Li, Shuang
Wang, Zhan-qi
Jiang, Tong
Functional analysis of a viral promoter from a strawberry vein banding virus isolate from China
title Functional analysis of a viral promoter from a strawberry vein banding virus isolate from China
title_full Functional analysis of a viral promoter from a strawberry vein banding virus isolate from China
title_fullStr Functional analysis of a viral promoter from a strawberry vein banding virus isolate from China
title_full_unstemmed Functional analysis of a viral promoter from a strawberry vein banding virus isolate from China
title_short Functional analysis of a viral promoter from a strawberry vein banding virus isolate from China
title_sort functional analysis of a viral promoter from a strawberry vein banding virus isolate from china
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8974135/
https://www.ncbi.nlm.nih.gov/pubmed/35361243
http://dx.doi.org/10.1186/s12985-022-01778-2
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