FADS1基因对猪不饱和脂肪酸含量的影响研究

doi:10.16431/j.cnki.1671-7236.2023.02.003

Abstract

Abstract: 【Objective】 To study the regulatory effect of fatty acid desaturases 1 (FADS1) gene on the metabolism of unsaturated fatty acids, and provide a reference for revealing its molecular mechanism. 【Method】 The FADS1 gene was overexpressed in the primary fetal fibroblasts isolated by vector pcDNA3.1-FLAG-FADS1, which was designed according to the CDS sequence of FADS1 gene. The constructed vector was transferred into the cells and screened by geneticin (G418) to obtain monoclonal cell lines. After transfection, cells were collected at 24 and 48 h respectively, and the expression of FADS1 gene was detected by Real-time PCR and western blotting, and the cell line with stable expression of FADS1 gene was obtained after G418 screening, named 1-4^#. These cells were used to detect the change of triglyceride content and analysis of unsaturated fatty acid composition through GC-MS.【Result】 The results showed that the overexpression vector pcDNA3.1-FLAG-FADS1 was successfully obtained, and the overexpression effect was detected at both the transcription level and the protein level. Then these cells were cultured in complete medium with G418 to obtain stable cell lines. Triglyceride assay showed that FADS1 gene overexpression could reduce the content of triglyceride in primary fetal fibroblasts (P＜0.05). The test results of unsaturated fatty acids showed that after overexpression of FADS1, the content of total fatty acids increased extremely significantly (P＜0.01) and increased by 1.8 times. In polyunsaturated fatty acids (PUFA), the contents of ω-3 PUFA and ω-6 PUFA were extremely significantly increased (P＜0.01), the relative contents of eicosapentaenoic acid(EPA), docosapentaenoic acid(DPA) and docosahexaenoic acid(DHA) were extremely increased (P＜0.01), and the ratio of ω-3/ω-6 was significantly increased (P＜0.05).【Conclusion】 In summary, the overexpression vector pcDNA 3.1-FLAG-FADS1 was successfully constructed, and a single clonal cell with stable hyperexpression of FADS1 gene was successfully screened in primary fetal fibroblasts, FADS1 gene could significantly increase the content of ω-3 PUFA and increase the ratio of ω-3/ω-6.

Key words: FADS1 gene; overexpression; fatty acids; ω-3; ω-6

CLC Number:

S828

ZHANG Tiantian, HUA Zaidong, REN Hongyan, GU Hao, ZHOU Bin, BI Yanzhen. Effect of FADS1 Gene on Unsaturated Fatty Acid Synthesis[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 451-459.

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Effect of FADS1 Gene on Unsaturated Fatty Acid Synthesis

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