Polymorphism Detection And Function Verifiction With Lipid Metabolism Of AGPAT6 Gene In Cattle

1-acylglycerol-3-phosphate-O-acyltransferases 6(AGPAT6) is one of the AGPATs family and it possesses GPAT activity. AGPAT6 is highly expressed in brown fat, mammary epithelial cell and many other tissues, and it has a major impact on diverse life activities.In the study, PCR-RFLP technique was used to test the genetic polymorphism of AGPAT6 gene. The association of the single nucleotide polymorphisms(SNPs) and the traits of dairy cattle and beef cattle was analyzed. Two SNPs were detected in Chinese Holstein steers. The polymorphism site E1-98A>G was not associated with traits of Chinese Holstein steers. And the polymorphism site E1-303T>C was significantly associated with the milk fat content, dry matter content, urea nitrogen content and milk yield(p<0.05). For Chinese Simmental steers, two SNPs were detected in the exon 1 and exon 12. The polymorphism site E1-303T>C was significantly associated with the omasum weight, liver weight, p H value of the beef, fat coverage rate of the carcass, fat color score, kidney weight, back fat thickness and heptadecenoic acid(p<0.05). And the polymorphism site E12-299G>A was significantly associated with the net weights of the bone, forehoof weight, hind feet weight, the fat coverage rate of the carcass, the head and heart weights and the marbling score(p<0.05).Bovine AGPAT6 overexpression vector and RNA interference vector were constructed. Bovine mammary epithelial cells and bovine fetal fibroblast cells were transfected by them separately and the indicators associated with lipid metabolism were tested. The results showed that bovine AGPAT6 could regulate m RNA expression levels of GPAT1, AGPAT1, DGAT1 and SREBP-1c gene in both two cells. At the same time, bovine AGPAT6 overexpression significantly increased triglyceride content and knockdown of AGPAT6 expression significantly reduced triglyceride content(p<0.05). AGPAT6 gene might play a important role in lipid metabolism.