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Construction and characterization of a synthetic Baculovirus-inducible 39K promoter

BACKGROUND: Silkworm genetic engineering is widely used in gene function, silk engineering and disease-resistant engineering in most of Asia. Some of the earliest promoter elements are used to control the development of silkworm transgenic expression and gene therapy. However, the low expression and...

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Autores principales: Dong, Zhan-Qi, Hu, Zhi-Gang, Li, Hai-Qing, Jiang, Ya-Ming, Cao, Ming-Ya, Chen, Peng, Lu, Cheng, Pan, Min-Hui
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6280533/
https://www.ncbi.nlm.nih.gov/pubmed/30534200
http://dx.doi.org/10.1186/s13036-018-0121-8
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author Dong, Zhan-Qi
Hu, Zhi-Gang
Li, Hai-Qing
Jiang, Ya-Ming
Cao, Ming-Ya
Chen, Peng
Lu, Cheng
Pan, Min-Hui
author_facet Dong, Zhan-Qi
Hu, Zhi-Gang
Li, Hai-Qing
Jiang, Ya-Ming
Cao, Ming-Ya
Chen, Peng
Lu, Cheng
Pan, Min-Hui
author_sort Dong, Zhan-Qi
collection PubMed
description BACKGROUND: Silkworm genetic engineering is widely used in gene function, silk engineering and disease-resistant engineering in most of Asia. Some of the earliest promoter elements are used to control the development of silkworm transgenic expression and gene therapy. However, the low expression and specificity of natural promoters limit the applications of genetic engineering. To construct a highly efficient synthetic inducible promoter in the Bombyx mori (Lepidoptera), we analyzed the regulatory elements and functional regions of the B. mori nucleopolyhedrovirus 39 K promoter. RESULTS: Truncated mutation analysis of the 39 K promoter showed that the transcriptional regulatory region spanning positions − 573 to − 274 and + 1 to + 62 are essential for virus-inducible promoter activity. Further investigations using the electrophoretic mobility shift assay revealed that the baculovirus IE-1 protein binds to the 39 K promoter at the − 310 to − 355 region, and transcription activates the expression of 39 K promoter assay. Finally, we successfully constructed a synthetic inducible promoter that increased the virus-inducing activity of other promoters using the baculovirus-inducible transcriptional activation region that binds to specific core elements of 39 K (i.e., spanning the region − 310 to − 355). CONCLUSIONS: In summary, we constructed a novel, synthetic, and highly efficient biological tool, namely, a virus-inducible 39 K promoter, which provides endless possibilities for future research on gene function, gene therapy, and pest control in genetic engineering. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13036-018-0121-8) contains supplementary material, which is available to authorized users.
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spelling pubmed-62805332018-12-10 Construction and characterization of a synthetic Baculovirus-inducible 39K promoter Dong, Zhan-Qi Hu, Zhi-Gang Li, Hai-Qing Jiang, Ya-Ming Cao, Ming-Ya Chen, Peng Lu, Cheng Pan, Min-Hui J Biol Eng Research BACKGROUND: Silkworm genetic engineering is widely used in gene function, silk engineering and disease-resistant engineering in most of Asia. Some of the earliest promoter elements are used to control the development of silkworm transgenic expression and gene therapy. However, the low expression and specificity of natural promoters limit the applications of genetic engineering. To construct a highly efficient synthetic inducible promoter in the Bombyx mori (Lepidoptera), we analyzed the regulatory elements and functional regions of the B. mori nucleopolyhedrovirus 39 K promoter. RESULTS: Truncated mutation analysis of the 39 K promoter showed that the transcriptional regulatory region spanning positions − 573 to − 274 and + 1 to + 62 are essential for virus-inducible promoter activity. Further investigations using the electrophoretic mobility shift assay revealed that the baculovirus IE-1 protein binds to the 39 K promoter at the − 310 to − 355 region, and transcription activates the expression of 39 K promoter assay. Finally, we successfully constructed a synthetic inducible promoter that increased the virus-inducing activity of other promoters using the baculovirus-inducible transcriptional activation region that binds to specific core elements of 39 K (i.e., spanning the region − 310 to − 355). CONCLUSIONS: In summary, we constructed a novel, synthetic, and highly efficient biological tool, namely, a virus-inducible 39 K promoter, which provides endless possibilities for future research on gene function, gene therapy, and pest control in genetic engineering. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13036-018-0121-8) contains supplementary material, which is available to authorized users. BioMed Central 2018-12-04 /pmc/articles/PMC6280533/ /pubmed/30534200 http://dx.doi.org/10.1186/s13036-018-0121-8 Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Dong, Zhan-Qi
Hu, Zhi-Gang
Li, Hai-Qing
Jiang, Ya-Ming
Cao, Ming-Ya
Chen, Peng
Lu, Cheng
Pan, Min-Hui
Construction and characterization of a synthetic Baculovirus-inducible 39K promoter
title Construction and characterization of a synthetic Baculovirus-inducible 39K promoter
title_full Construction and characterization of a synthetic Baculovirus-inducible 39K promoter
title_fullStr Construction and characterization of a synthetic Baculovirus-inducible 39K promoter
title_full_unstemmed Construction and characterization of a synthetic Baculovirus-inducible 39K promoter
title_short Construction and characterization of a synthetic Baculovirus-inducible 39K promoter
title_sort construction and characterization of a synthetic baculovirus-inducible 39k promoter
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6280533/
https://www.ncbi.nlm.nih.gov/pubmed/30534200
http://dx.doi.org/10.1186/s13036-018-0121-8
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