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| Cloning of Elongase of Very Long Chain Fatty Acids 6 Gene(ELOVL6)and Its Effect on Four Genes Expression Involved in Fatty Acid Metabolism in Dairy Goat(Capra hircus) |
| 1,Jun LUO 3, 3, 3, 3, 3 |
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Abstract Elongase of very long chain fatty acids 6 (ELOVL6) is the rate-limiting enzyme in the elongation cycle in mammals. In this study, ELOVL6 coding sequences (CDS) was cloned by RT-PCR, tissue expession was analyzed by Real-time quantitative PCR (qRT-PCR) and constructed a recombinant Adenovirus overexpression vector for ELOVL6 of dairy goat (Capra hircus) to investigate its effect of related genes on fatty acid metabolism after infecting recombinant Adenovirus in dairy goat mammary epithelial cells(GMEC). The results showed that the CDS of ELOVL6 gene was 795 bp (GenBank No. KF667508), encoding 264 amino acids. Compared with cattle(Bos taurus), rat(Rattus norvegicus) and human (Homo sapiens), the nucleic acid sequence homology of dairy goat ELOVL6 was 97%, 90% and 93%, and the amino acid sequence homology was 99%, 92% and 95%, respectively. The protein structure analysis using TMpred showed that 5 transmembrane helixes were speculated in ELOVL6 protein, and there was a conserved histidine box (HWYHH) between the second and the third transmembrane helix. The ELOVL6 protein had strong hydrophobic properties analysed by ProtScale. Tissue expression analysis showed that ELOVL6 gene was themost abundant in adipose tissue(P<0.01), followed by small intestine(P<0.01), and minimum in heart. The titer of recombinant Adenovirus containing ELOVL6 gene was 108 U/mL. Compared with Ad-GFP group, ELOVL6 mRNA level increased by about 200-fold in in GMEC after incubated with the virus for 48 h. Sterol regulatory element binding protein-1 (SREBP-1) was decreased significantly (P<0.05), and adipose differentiation related protein (ADRP) was increased significantly (P<0.05). There was no significant effect on the expression of peroxisome proliferator activated receptor γ (PPARγ) and fatty acid translocase (CD36). These results indicated that ELOVL6 can influence the expression of genes related to fatty acid metabolism, and may have certain function on fatty acid metabolism in dairy goat mammary epithelial cells. This study provides the theoretical foundation for the study of fatty acid metabolism in the process of lactation.
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Received: 16 December 2013
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Corresponding Authors:
Jun LUO
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