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Cloning and Molecular Characterization of Porcine β-casein Gene (CNS2)

The production of therapeutic proteins from transgenic animals is one of the most important successes of animal biotechnology. Milk is presently the most mature system for production of therapeutic proteins from a transgenic animal. Specifically, β-casein is a major component of cow, goat and sheep...

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Autores principales: Lee, Sang Mi, Kim, Hye-Min, Moon, Seung Ju, Kang, Man-Jong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Asian-Australasian Association of Animal Production Societies (AAAP) and Korean Society of Animal Science and Technology (KSAST) 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4092958/
https://www.ncbi.nlm.nih.gov/pubmed/25049581
http://dx.doi.org/10.5713/ajas.2011.11240
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author Lee, Sang Mi
Kim, Hye-Min
Moon, Seung Ju
Kang, Man-Jong
author_facet Lee, Sang Mi
Kim, Hye-Min
Moon, Seung Ju
Kang, Man-Jong
author_sort Lee, Sang Mi
collection PubMed
description The production of therapeutic proteins from transgenic animals is one of the most important successes of animal biotechnology. Milk is presently the most mature system for production of therapeutic proteins from a transgenic animal. Specifically, β-casein is a major component of cow, goat and sheep milk, and its promoter has been used to regulate the expression of transgenic genes in the mammary gland of transgenic animals. Here, we cloned the porcine β-casein gene and analyzed the transcriptional activity of the promoter and intron 1 region of the porcine β-casein gene. Sequence inspection of the 5′-flanking region revealed potential DNA elements including SRY, CdxA, AML-a, GATA-3, GATA-1 and C/EBP β. In addition, the first intron of the porcine β-casein gene contained the transcriptional enhancers Oct-1, SRY, YY1, C/EBP β, and AP-1, as well as the retroviral TATA box. We estimated the transcriptional activity for the 5′-proximal region with or without intron 1 of the porcine β-casein gene in HC11 cells stimulated with lactogenic hormones. High transcriptional activity was obtained for the 5′-proximal region with intron 1 of the porcine β-casein gene. The β-casein gene containing the mutant TATA box (CATAAAA) was also cloned from another individual pig. Promoter activity of the luciferase vector containing the mutant TATA box was weaker than the same vector containing the normal TATA box. Taken together, these findings suggest that the transcription of porcine β-casein gene is regulated by lactogenic hormone via intron 1 and promoter containing a mutant TATA box (CATAAAA) has poor porcine β-casein gene activity.
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spelling pubmed-40929582014-07-21 Cloning and Molecular Characterization of Porcine β-casein Gene (CNS2) Lee, Sang Mi Kim, Hye-Min Moon, Seung Ju Kang, Man-Jong Asian-Australas J Anim Sci Article The production of therapeutic proteins from transgenic animals is one of the most important successes of animal biotechnology. Milk is presently the most mature system for production of therapeutic proteins from a transgenic animal. Specifically, β-casein is a major component of cow, goat and sheep milk, and its promoter has been used to regulate the expression of transgenic genes in the mammary gland of transgenic animals. Here, we cloned the porcine β-casein gene and analyzed the transcriptional activity of the promoter and intron 1 region of the porcine β-casein gene. Sequence inspection of the 5′-flanking region revealed potential DNA elements including SRY, CdxA, AML-a, GATA-3, GATA-1 and C/EBP β. In addition, the first intron of the porcine β-casein gene contained the transcriptional enhancers Oct-1, SRY, YY1, C/EBP β, and AP-1, as well as the retroviral TATA box. We estimated the transcriptional activity for the 5′-proximal region with or without intron 1 of the porcine β-casein gene in HC11 cells stimulated with lactogenic hormones. High transcriptional activity was obtained for the 5′-proximal region with intron 1 of the porcine β-casein gene. The β-casein gene containing the mutant TATA box (CATAAAA) was also cloned from another individual pig. Promoter activity of the luciferase vector containing the mutant TATA box was weaker than the same vector containing the normal TATA box. Taken together, these findings suggest that the transcription of porcine β-casein gene is regulated by lactogenic hormone via intron 1 and promoter containing a mutant TATA box (CATAAAA) has poor porcine β-casein gene activity. Asian-Australasian Association of Animal Production Societies (AAAP) and Korean Society of Animal Science and Technology (KSAST) 2012-03 /pmc/articles/PMC4092958/ /pubmed/25049581 http://dx.doi.org/10.5713/ajas.2011.11240 Text en Copyright © 2012 by Asian-Australasian Journal of Animal Sciences This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License http://creativecommons.org/licenses/by-nc/3.0/ which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Article
Lee, Sang Mi
Kim, Hye-Min
Moon, Seung Ju
Kang, Man-Jong
Cloning and Molecular Characterization of Porcine β-casein Gene (CNS2)
title Cloning and Molecular Characterization of Porcine β-casein Gene (CNS2)
title_full Cloning and Molecular Characterization of Porcine β-casein Gene (CNS2)
title_fullStr Cloning and Molecular Characterization of Porcine β-casein Gene (CNS2)
title_full_unstemmed Cloning and Molecular Characterization of Porcine β-casein Gene (CNS2)
title_short Cloning and Molecular Characterization of Porcine β-casein Gene (CNS2)
title_sort cloning and molecular characterization of porcine β-casein gene (cns2)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4092958/
https://www.ncbi.nlm.nih.gov/pubmed/25049581
http://dx.doi.org/10.5713/ajas.2011.11240
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