Molecular Cloning of a Functional Fads2 Promoter from Zebrafish

The FADS2 catalyzes the first rate-limiting step in the long chain-polyunsaturated fatty acids (LC-PUFAs) biosynthesis pathway by converting �-linolenic acid and linoleic acid into stearidonic acid and �-linolenic acid via the �-3 and �-6 pathways respectively. In mammals, PPAR� and SREBP-1c have...

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Main Authors: Chung, Hung Hui, Azham, Zulkharnain
Format: Article
Language:English
Published: Hibiscus Publisher 2016
Subjects:
Online Access:http://ir.unimas.my/id/eprint/13689/
http://ir.unimas.my/id/eprint/13689/1/Molecular%20Cloning.pdf
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author Chung, Hung Hui
Azham, Zulkharnain
author_facet Chung, Hung Hui
Azham, Zulkharnain
author_sort Chung, Hung Hui
building UNIMAS Institutional Repository
collection Online Access
description The FADS2 catalyzes the first rate-limiting step in the long chain-polyunsaturated fatty acids (LC-PUFAs) biosynthesis pathway by converting �-linolenic acid and linoleic acid into stearidonic acid and �-linolenic acid via the �-3 and �-6 pathways respectively. In mammals, PPAR� and SREBP-1c have been implicated in the polyunsaturated fatty acids (PUFAs) mediated transcriptional activation of FADS2 promoter. However, in zebrafish, not much is known regarding the regulation of fads2 transcriptional regulation. Here, in this study, five vectors containing different promoter regions were constructed in order to analyse putative promoter activities. Through truncation analysis, it was found that the 1.2 kb promoter was able to drive luciferase activity to an approximate 40-fold in HepG2 cells. Upon mutagenesis analysis, three sites which are the putative NF-Y, SREBP and PPAR binding sites were found to be essential in driving the promoter activity. Lastly, the 1.2 kb fads2 promoter was able to direct EGFP expression specifically to the yolk syncytial layer (YSL) when transiently expressed in microinjected zebrafish embryos.
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spelling unimas-136892022-02-04T07:28:14Z http://ir.unimas.my/id/eprint/13689/ Molecular Cloning of a Functional Fads2 Promoter from Zebrafish Chung, Hung Hui Azham, Zulkharnain SH Aquaculture. Fisheries. Angling The FADS2 catalyzes the first rate-limiting step in the long chain-polyunsaturated fatty acids (LC-PUFAs) biosynthesis pathway by converting �-linolenic acid and linoleic acid into stearidonic acid and �-linolenic acid via the �-3 and �-6 pathways respectively. In mammals, PPAR� and SREBP-1c have been implicated in the polyunsaturated fatty acids (PUFAs) mediated transcriptional activation of FADS2 promoter. However, in zebrafish, not much is known regarding the regulation of fads2 transcriptional regulation. Here, in this study, five vectors containing different promoter regions were constructed in order to analyse putative promoter activities. Through truncation analysis, it was found that the 1.2 kb promoter was able to drive luciferase activity to an approximate 40-fold in HepG2 cells. Upon mutagenesis analysis, three sites which are the putative NF-Y, SREBP and PPAR binding sites were found to be essential in driving the promoter activity. Lastly, the 1.2 kb fads2 promoter was able to direct EGFP expression specifically to the yolk syncytial layer (YSL) when transiently expressed in microinjected zebrafish embryos. Hibiscus Publisher 2016 Article PeerReviewed text en http://ir.unimas.my/id/eprint/13689/1/Molecular%20Cloning.pdf Chung, Hung Hui and Azham, Zulkharnain (2016) Molecular Cloning of a Functional Fads2 Promoter from Zebrafish. Journal of biochemistry, microbiology and biotechnology, 4 (1). pp. 1-6. ISSN 2289-5779 http://journal.hibiscuspublisher.com/index.php/JOBIMB/article/view/280/326
spellingShingle SH Aquaculture. Fisheries. Angling
Chung, Hung Hui
Azham, Zulkharnain
Molecular Cloning of a Functional Fads2 Promoter from Zebrafish
title Molecular Cloning of a Functional Fads2 Promoter from Zebrafish
title_full Molecular Cloning of a Functional Fads2 Promoter from Zebrafish
title_fullStr Molecular Cloning of a Functional Fads2 Promoter from Zebrafish
title_full_unstemmed Molecular Cloning of a Functional Fads2 Promoter from Zebrafish
title_short Molecular Cloning of a Functional Fads2 Promoter from Zebrafish
title_sort molecular cloning of a functional fads2 promoter from zebrafish
topic SH Aquaculture. Fisheries. Angling
url http://ir.unimas.my/id/eprint/13689/
http://ir.unimas.my/id/eprint/13689/
http://ir.unimas.my/id/eprint/13689/1/Molecular%20Cloning.pdf