The functional analysis of the ocean pout (Macrozoarces americanus) type III antifreeze protein gene promoter
The full length ocean pout antifreeze gene promoter and a series of promoter deletion constructs were fused to the luciferase reporter gene, and their transcriptional role was examined in a transient expression assay by transfection into three cell lines; a human epithelial (CRL-1578), a human fibro...
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Memorial University of Newfoundland
2005
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ftmemorialuniv:oai:research.library.mun.ca:10814 2023-10-01T03:54:47+02:00 The functional analysis of the ocean pout (Macrozoarces americanus) type III antifreeze protein gene promoter Kirby, Trina Maxine 2005 application/pdf https://research.library.mun.ca/10814/ https://research.library.mun.ca/10814/1/Kirby_TrinaMaxine.pdf en eng Memorial University of Newfoundland https://research.library.mun.ca/10814/1/Kirby_TrinaMaxine.pdf Kirby, Trina Maxine <https://research.library.mun.ca/view/creator_az/Kirby=3ATrina_Maxine=3A=3A.html> (2005) The functional analysis of the ocean pout (Macrozoarces americanus) type III antifreeze protein gene promoter. Masters thesis, Memorial University of Newfoundland. thesis_license Thesis NonPeerReviewed 2005 ftmemorialuniv 2023-09-03T06:48:01Z The full length ocean pout antifreeze gene promoter and a series of promoter deletion constructs were fused to the luciferase reporter gene, and their transcriptional role was examined in a transient expression assay by transfection into three cell lines; a human epithelial (CRL-1578), a human fibroblast (HTB-92) and a Chinook salmon embryonic cell line (CRL-1691). The studies demonstrate that the ocean pout AFP gene promoter can function in both mammalian and fish cell lines and that they contain both positive as well as negative regulatory regions. This indicates that the AFP gene may be regulated at various sites. -- In previous studies, a fish antifreeze/ growth hormone chimeric gene construct opAFP-GHc2 was injected into Atlantic salmon eggs. Through PCR analysis it was found that this construct reorganized in the fish genome so that the beginning of the 5’ ocean pout AFP promoter end was linked to the 3' end of the AFP polyadenylation sequence. In my study, this reorganized transgenic promoter was also ligated to the luciferase reporter gene and examined in a transient transfection assay, which determined that this reorganized promoter had significantly less activity than that of the full-length intact promoter. However, promoter activity was detected in all three cell lines which indicates that the transgene should be expected to express growth hormone in vivo. Thesis Atlantic salmon Memorial University of Newfoundland: Research Repository |
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Memorial University of Newfoundland: Research Repository |
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ftmemorialuniv |
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English |
description |
The full length ocean pout antifreeze gene promoter and a series of promoter deletion constructs were fused to the luciferase reporter gene, and their transcriptional role was examined in a transient expression assay by transfection into three cell lines; a human epithelial (CRL-1578), a human fibroblast (HTB-92) and a Chinook salmon embryonic cell line (CRL-1691). The studies demonstrate that the ocean pout AFP gene promoter can function in both mammalian and fish cell lines and that they contain both positive as well as negative regulatory regions. This indicates that the AFP gene may be regulated at various sites. -- In previous studies, a fish antifreeze/ growth hormone chimeric gene construct opAFP-GHc2 was injected into Atlantic salmon eggs. Through PCR analysis it was found that this construct reorganized in the fish genome so that the beginning of the 5’ ocean pout AFP promoter end was linked to the 3' end of the AFP polyadenylation sequence. In my study, this reorganized transgenic promoter was also ligated to the luciferase reporter gene and examined in a transient transfection assay, which determined that this reorganized promoter had significantly less activity than that of the full-length intact promoter. However, promoter activity was detected in all three cell lines which indicates that the transgene should be expected to express growth hormone in vivo. |
format |
Thesis |
author |
Kirby, Trina Maxine |
spellingShingle |
Kirby, Trina Maxine The functional analysis of the ocean pout (Macrozoarces americanus) type III antifreeze protein gene promoter |
author_facet |
Kirby, Trina Maxine |
author_sort |
Kirby, Trina Maxine |
title |
The functional analysis of the ocean pout (Macrozoarces americanus) type III antifreeze protein gene promoter |
title_short |
The functional analysis of the ocean pout (Macrozoarces americanus) type III antifreeze protein gene promoter |
title_full |
The functional analysis of the ocean pout (Macrozoarces americanus) type III antifreeze protein gene promoter |
title_fullStr |
The functional analysis of the ocean pout (Macrozoarces americanus) type III antifreeze protein gene promoter |
title_full_unstemmed |
The functional analysis of the ocean pout (Macrozoarces americanus) type III antifreeze protein gene promoter |
title_sort |
functional analysis of the ocean pout (macrozoarces americanus) type iii antifreeze protein gene promoter |
publisher |
Memorial University of Newfoundland |
publishDate |
2005 |
url |
https://research.library.mun.ca/10814/ https://research.library.mun.ca/10814/1/Kirby_TrinaMaxine.pdf |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_relation |
https://research.library.mun.ca/10814/1/Kirby_TrinaMaxine.pdf Kirby, Trina Maxine <https://research.library.mun.ca/view/creator_az/Kirby=3ATrina_Maxine=3A=3A.html> (2005) The functional analysis of the ocean pout (Macrozoarces americanus) type III antifreeze protein gene promoter. Masters thesis, Memorial University of Newfoundland. |
op_rights |
thesis_license |
_version_ |
1778522713708560384 |