奶山羊SCD基因CDS区的克隆、序列分析及过表达

doi:10.3864/j.issn.0578-1752.2012.24.015

Abstract

Abstract: 【Objective】To study the function of stearoyl-CoA desaturase (SCD) gene in mammary gland fatty acid metabolism emphasizing on how it regulates genes involved in unsaturated fatty acid synthesis as well as its impact on fatty acid composition.【Method】The sequence of SCD in dairy goat was cloned using RT-PCR for bioinformatics and tissue mRNA expression analysis. A recombinant adenovirus vector was constructed to over-express the gene in cultured goat mammary epithelial cells. Changes of gene expression, protein yield and fatty acid contents after over-expression were detected with real-time fluorescence quantitative PCR, western blotting and GC-MS. 【Result】The CDS of goat SCD gene is 1080 bp in length, coding 359 amino acid residues (GenBank accession: GU947654). The SCD gene was expressed higher in mammary gland, lung and subcutaneous fat, lower in heart, rumen and ovary. Recombinant adenovirus vector containing goat SCD open reading frame was constructed, packaged and amplified in HEK293. The gene expressions of FASN, A-FABP, LEPR and LXRα were down regulated after over-expression of SCD, while expressions of PPARγ and LPL were up regulated significantly. Synthesis of palmitic acid and stearic acid in the cells decreased, while unsaturated fatty acids increased.【Conclusion】 SCD gene plays an important role in goat mammary fatty acid metabolism, and a major role in unsaturated fatty acid synthesis by interaction with other genes including FASN, H-FABP, LEPR, LXRα, PPARγ and LPL.

Key words: dairy goat , stearoyl-CoA desaturase , mammary epithelial cells , adenovirus , over-expression

SHI Heng-Bo, LUO Jun, ZHU Yue, WANG Zi-Qian, ZHAO Wang-Sheng. Cloning, Sequence Analysis and Over-Expression of SCD Gene of Dairy Goat[J].Scientia Agricultura Sinica, 2012, 45(24): 5091-5101.

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Cloning, Sequence Analysis and Over-Expression of SCD Gene of Dairy Goat

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