Abstract
Intramuscular fat (IMF) content has been generally recognized as a desirable trait in pork meat because of its positive effect on eating quality. An effective approach to enhance IMF content in pork is the generation of transgenic pigs. In this study, we used somatic cell nuclear transfer (SCNT) to generate cloned pigs exhibiting ectopic expression of phosphoenolpyruvate carboxykinase (PEPCK-C) driven by an α-skeletal-actin gene promoter, which was specifically expressed in skeletal muscle. Using qRT-PCR and Western blot analysis, we demonstrated that PEPCK-C was functionally expressed and had a significant effect on total fatty acid content in the skeletal muscle of the transgenic pigs, while the n-6/n-3 polyunsaturated fatty acid (PUFA) ratio showed no difference between transgenic and control pigs. Thus, genetically engineered PEPCK-Cmus pigs may be an effective solution for the production of IMF-enriched pork.
Highlights
The cytosolic form of phosphoenolpyruvate carboxykinase (PEPCK-C) is a major rate-limiting enzyme in gluconeogenesis in the liver and kidney cortex and in glyceroneogenesis in the liver and white and brown adipose tissue[23]
Six (T01, T03, T04, T05, T09, T11) harbored the PEPCK-C transgene, as determined by PCR screening (Fig. 1c). These results indicate that six PEPCK-Cmus transgenic founder pigs were produced in this study
The skeletal muscle of PEPCK-Cmus transgenic pigs had a higher triglyceride content compared to that of control animals, consistent with the skeletal muscle phenotype first observed in PEPCK-Cmus mice[24]
Summary
The cytosolic form of phosphoenolpyruvate carboxykinase (PEPCK-C) is a major rate-limiting enzyme in gluconeogenesis in the liver and kidney cortex and in glyceroneogenesis in the liver and white and brown adipose tissue[23]. The metabolic role of this enzyme in other mammalian tissues remains unclear. Hakimi et al.[24] demonstrated that overexpression of the PEPCK-C gene in the skeletal muscle of mice increased triglyceride levels in skeletal muscle and decreased triglyceride levels in adipose tissue. This surprising metabolic outcome in mice prompted researchers to evaluate the developmental effects of the PEPCK-C transgene in livestock. The objectives of this study were to use somatic cell nuclear transfer (SCNT) to generate transgenic pigs carrying a PEPCK-C transgene regulated by the α-skeletal-actin gene promoter and to determine the effect of the PEPCK-C gene on the IMF content of the transgenic pigs
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