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| Construction of Interference Vector of FGF10 Gene and Expression Analysis of Related Genes in Congjiang Xiang Pig (Sus scrofa) |
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Abstract The fibroblast growth factor 10 gene (FGF10) is a member of the fibroblast growth factor family, it plays an important role in the development and metabolism of white adipose tissue. In order to further reveal the role of FGF10 gene in the regulation of intramuscular fat in Congjiang Xiang pigs (Sus scrofa), in this experiment, four pairs of siRNA interference sequences and a pair of negative control sequences were designed by online software,and were connected to pGPU6/GFP/Neo vector. The successfully constructed interference vectors were transfected into Congjiang Xiang pig intramuscular preadipocytes and the best interference vector was screened by qRT-PCR. The expression of FGF10 gene was inhibited by the best interference vector. And then, the expressions of peroxisome proliferator activated receptorγ (PPARγ), adipose triglyceride lipase (ATGL),adiponectin (ADIPOQ), fatty acid binding protein 4 (FABP4), fatty acid synthase (FAS), lipoprotein lipase(LPL), acetyl-CoA carboxylase (ACC) and Hormone-sensitive lipase (HSL) gene were detected by qRT-PCR.. The results showed that the interference vector of the FGF10 gene was constructed successfully, and the optimal interference vector was screened with the interference efficiency of 74.30%. After inhibiting the expression of FGF10 gene, the expression of ACC, ATGL, ADIPOQ, FABP4, FAS, LPL, PPARγ and HSL were decreased. The expression of ACC and PPARγ genes were significantly lower than the control group (P＜0.05), and the expression of ATGL, ADIPOQ, FABP4, FAS and LPL genes were significantly lower than control group (P＜0.01). There was no significant difference between the relative expression of HSL gene and the control group (P＞0.05). In this experiment, the constructed interference vector of FGF10 gene was successfully transfected into the intramuscular preadipocyte cells of Congjiang Xiang pig, and the procine intramuscular preadipocytes differentiation can be inhibited by sliencing of FGF10 gene in vitro, which might be related to down-regulation of fat differentiation genes. This study provides foundation for further research on the regulation mechanism of FGF10 gene on fatty deposits.
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Received: 14 May 2018
Published: 20 November 2018
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