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Swine PPAR-gamma 2 expression upregulated in skeletal muscle of transgenic mice via the swine Myozenin-1 gene promoter

机译:猪Myozenin-1基因启动子在转基因小鼠骨骼肌中的猪PPAR-γ2表达上调

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摘要

Myozenin-1 (Myoz1) gene-encoded calsarcin-2 protein was expressed exclusively in fast-twitch muscles. Peroxisome proliferator-activated receptor gamma 2 (PPAR-gamma 2) is a key regulator of adipocyte differentiation, fatty acid uptake and storage in mammals. In this study, transgenic (TG) mice were generated by injecting linearized DNA that contained mouse creatine kinase M-type enhancer, Myoz1 core promoter, swine PPAR-gamma 2 (sPPAR-gamma 2) and SV40 polyadenylation sequences into pronuclei of fertilized FVB/NJ mouse embryos using microinjection technology. Then, the TG mice were used to identify whether swine Myoz1 (sMyoz1) promoter could upregulate sPPAR-gamma 2 expression in skeletal muscle in a TG mouse model. The results showed that the sMyoz1 promoter indeed upregulated sPPAR-gamma 2 expression on both the RNA and protein levels. The target genes of PPAR-gamma in fat formation pathways, such as fatty acid-binding protein 4 (FABP4) and lipoprotein lipase (LPL), were also overexpressed on the RNA level. Meanwhile, the level of skeletal muscle triacylglycerol in TG mice was increased (P 0.05), and the result of Oil Red-O staining in the skeletal muscle sections also showed that the number of lipid droplets was significantly increased in TG mice compared to wild-type mice, which might improve the intramuscular fat (IMF) content. For pork, the quality was mostly influenced by the IMF; the identification of swine muscle-specific promoter, sMyoz1, could further serve to develop transgenic pigs with higher intramuscular fat contents and improve pork quality.
机译:Myozenin-1(Myoz1)基因编码的calsarcin-2蛋白仅在快肌中表达。过氧化物酶体增殖物激活受体γ2(PPAR-γ2)是哺乳动物脂肪细胞分化,脂肪酸摄取和储存的关键调节剂。在这项研究中,转基因(TG)小鼠是通过将线性化的DNA注入受精的FVB //使用显微注射技术的NJ小鼠胚胎。然后,将TG小鼠用于鉴定猪Myoz1(sMyoz1)启动子是否可以上调TG小鼠模型中骨骼肌中sPPAR-γ2的表达。结果表明,sMyoz1启动子确实在RNA和蛋白质水平上均上调了sPPAR-γ2表达。脂肪形成途径中PPAR-γ的靶基因,例如脂肪酸结合蛋白4(FABP4)和脂蛋白脂肪酶(LPL),也在RNA水平上过表达。同时,TG小鼠的骨骼肌三酰甘油水平增加(P <0.05),骨骼肌切片上的油红-O染色结果也表明,TG小鼠的脂滴数量比野生动物显着增加型小鼠,可能会改善肌内脂肪(IMF)含量。对于猪肉,质量主要受IMF影响。猪肌肉特异性启动子sMyoz1的鉴定可以进一步开发出具有较高肌内脂肪含量的转基因猪,并改善猪肉品质。

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