Construction Of FDFT1 ShRNA And Overexpression Vector And Effects Cholesterol Metabolism In Bovine Fetal Fibroblast

FDFT1 gene(Farnesyl-Diphosphate Farnesyltransferase 1)is an important gene in the metabolic pathway of cholesterol.To understand the function of FDFT1 in cholesterol metabolism and lipid metabolism,overexpression vectors of FDFT1 gene and shRNA vector with green fluorescent protein(GFP)were constructed which transferred into Bovine Fetal Fibroblasts(BFF)and detected the expression levels of mRNA and protein.After using the successful construction of carrier interference or overexpression of FDFT1 gene,FDFT1 gene is located on the downstream pathway of cholesterol biosynthesis,metabolism and expression analysis of cholesterol cholesterol transport related genes,further reveals the interactions of FDFT1 gene synthesis in cholesterol metabolism and transport.By testing the content of cholesterol and triglycerides also can understand the role of FDFT1 gene in lipid metabolism.The result showed that compared with shRNA NC group expression levels of mRNA in shRNA group were downregulated 40.7 %(p<0.01)and protein were downregulated 39.61%(p<0.01);in pBI-CMV3-FDFT1 overexpression group expression levels of mRNA were upregulated 137.9 times than pBI-CMV3(p<0.01)and protein were only 2.34 times(p<0.01).This study detected the mRNA and protein expression levels by RT-PCR and western blot after transfection to verify the effectiveness of shRNA and overexpression vectors of bovine FDFT1 gene and provided an molecular basis for further study of FDFT1 in cholesterol metabolism and lipid metabolism.The interference or overexpression of FDFT1 gene changes its expression,which eventually can cause the passage of HMGCR,SQLE,MVD,DHCR7,NSDHL,FDPS,SC5 DL,CYP51A1,LSS and HMGCS1;cholesterol biosynthesis CNBP,DHCR24,EBP,PRKAA2,PRKAG2;cholesterol metabolism CYP46A1,SCARF1,TRERF1,CYP7B1,CELA3 B,HDLBP,APOB and LDLRAP1 and cholesterol transport ABCG1,APOA1 and STARD3 expression change.The interference making the FDFT1 gene expression decreased also cause significantly decreased the contents of triglyceride and cholesterol,while the overexpression of FDFT1 gene by increasing the expression makes the triglyceride and cholesterol expression significantly increased(p<0.01).