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Functional expression of a humanized gene for an ω-3 fatty acid desaturase from scarlet flax in transfected bovine adipocytes and bovine embryos cloned from the cells
- Source :
- Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids. 1791:183-190
- Publication Year :
- 2009
- Publisher :
- Elsevier BV, 2009.
-
Abstract
- Long-chain n-3 fatty acids can lower the risk of lifestyle-related diseases, therefore, we introduced a plant fatty acid desaturation3 (FAD3) gene into mammalian cells. The FAD3 cDNA was isolated from the immature seeds of scarlet flax and optimized to human high-frequency codon usage for enhancement of its expression levels in mammalian cells (hFAD3). We introduced the gene into bovine muscle satellite cells, which can be differentiated into multilocular adipocytes in vitro. After hFAD3 transfection, the cells were differentiated into adipocytes and their fatty acid composition was analyzed by gas chromatography. The level of alpha-linolenic acid (18:3n-3) in transfected adipocytes increased about ten-fold compared with non-transfected adipocytes. In addition, the levels of docosapentaenoic acid (DPA, 22:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) in transfected adipocytes were significantly higher than those in non-transfected adipocytes. Moreover, we produced bovine cloned embryos from the hFAD3 cells by somatic cell nuclear transfer. Blastocyst rates of hFAD3 clones were the same as the control clones using the non-transfected cells (21% vs 27%, P > 0.05). hFAD3 transcripts were detected in all of the blastocysts. These results demonstrate the functional expression of a plant hFAD3 in mammalian adipocytes, and normal development of cloned embryos carrying the hFAD3 gene.
- Subjects :
- Fatty Acid Desaturases
Male
Chromatography, Gas
DNA, Complementary
Docosahexaenoic Acids
Satellite Cells, Skeletal Muscle
Cellular differentiation
Transfection
Gene Expression Regulation, Enzymologic
Embryo Culture Techniques
chemistry.chemical_compound
Flax
Gene expression
Adipocytes
Animals
Humans
Molecular Biology
Cells, Cultured
chemistry.chemical_classification
biology
alpha-Linolenic acid
fungi
alpha-Linolenic Acid
Fatty acid
Cell Biology
Embryo, Mammalian
Molecular biology
Blastocyst
Fatty acid desaturase
chemistry
Docosahexaenoic acid
embryonic structures
Fatty Acids, Unsaturated
biology.protein
Cattle
Docosapentaenoic acid
Subjects
Details
- ISSN :
- 13881981
- Volume :
- 1791
- Database :
- OpenAIRE
- Journal :
- Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids
- Accession number :
- edsair.doi.dedup.....6ed062261deab95396eacb2b1af6204f
- Full Text :
- https://doi.org/10.1016/j.bbalip.2008.12.019