Transgenic technology has been applied to generate new high value-added livestock animals to meet the growing demand for healthy food. So far, FAD3 derived from spinach, Caenorhabditis elegans (C. elegans) andCaenorhabditis briggsae (C. briggsae) have been heterologously expressed in pigs (Zhou et al. 2014; Pan etal. 2010; Laietal. 2006; Saekietal. 2004), dairy cattle (Wu et al. 2012) and sheep (Zhang et al. 2013). C. elegans fat1 gene, which encodes FAD3, is a good candidate gene to breed into new transgenic animals to increase the contents of n-3 PUFAs. To the best of our knowledge, no fat1 transgenic beef cattle have been obtained up to now. Therefore, in this study, mfat1 (codon-optimized C. elegans fat1 gene) transgenic beef cattle were first generated via somatic cell nuclear transfer (SCNT). A plasmid containing the codon-optimized C. elegans mfat1 gene was constructed and transfected into the primary fetal fibroblasts from Luxi Yellow cattle. Luxi Yellow is a well-known Chinese breed of indigenous cattle, with a high beef yield. The obtained mfat1 transgenic Luxi cattle heterologously expressed FAD3, and showed significantly increased contents of n-3 PUFAs in muscle tissue. Also, the ratio of n-6/n-3 PUFAs in transgenic cattle was 0.95: 1, which was lower than that 5.33: 1 of transgenic negative cattle. Our results demonstrate that the mfat1 gene from C. elegans could be functionally expressed in beef cattle and will decrease the ratio of n-6/n-3 PUFAs in muscle tissue, which may make beef a good source of n-3 PUFAs in human daily diet.
Transgenic technology has been applied to generate new high value-added livestock animals to meet the growing demand for healthy food. So far, FAD3 derived from spinach, Caenorhabditis elegans (C. elegans) andCaenorhabditis briggsae (C. briggsae) have been heterologously expressed in pigs (Zhou et al. 2014; Pan etal. 2010; Laietal. 2006; Saekietal. 2004), dairy cattle (Wu et al. 2012) and sheep (Zhang et al. 2013). C. elegans fat1 gene, which encodes FAD3, is a good candidate gene to breed into new transgenic animals to increase the contents of n-3 PUFAs. To the best of our knowledge, no fat1 transgenic beef cattle have been obtained up to now. Therefore, in this study, mfat1 (codon-optimized C. elegans fat1 gene) transgenic beef cattle were first generated via somatic cell nuclear transfer (SCNT). A plasmid containing the codon-optimized C. elegans mfat1 gene was constructed and transfected into the primary fetal fibroblasts from Luxi Yellow cattle. Luxi Yellow is a well-known Chinese breed of indigenous cattle, with a high beef yield. The obtained mfat1 transgenic Luxi cattle heterologously expressed FAD3, and showed significantly increased contents of n-3 PUFAs in muscle tissue. Also, the ratio of n-6/n-3 PUFAs in transgenic cattle was 0.95: 1, which was lower than that 5.33: 1 of transgenic negative cattle. Our results demonstrate that the mfat1 gene from C. elegans could be functionally expressed in beef cattle and will decrease the ratio of n-6/n-3 PUFAs in muscle tissue, which may make beef a good source of n-3 PUFAs in human daily diet.
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