Construction of Recombinant Eukaryotic Expression Vector of FoxO4 Gene from Congjiang Xiang Pig and Its Expression in Subcutaneous Preadipocytes
The aim of this study was to clone the coding sequence of FoxO 4 gene of Congjiang Xiang pig and preliminarily explore the function of FoxO 4 gene.The relative expression level of FoxO 4 gene was detected by qRT-PCR in different tissues of Congjiang Xiang pig at the age of 6 month.The recombinant plasmid pEGFP-N3-FoxO4 was constructed by inserting the FoxO 4 gene coding sequence into pEGFP-N3,and then transiently transfected into the subcutaneous preadipocytes of Congjiang Xiang pig by liposome-mediated transfection.The expression levels of FoxO 4,PPARγ,LPL,FAS,ACC,ATGL,HSL in transfected cells were detected by qRT-PCR.The results showed that FoxO 4 gene had the highest expression level in adipose tissue.The constructed plasmid pEGFP-N3-FoxO4 was successfully expressed in subcutaneous preadipocytes.The expression level of LPL,FAS,ACC,ATGL and HSL genes were significantly upregulated in the subcutaneous preadipocytes when compared to the control group.These results revealed that FoxO 4 gene had certain regulation on fat deposition and might be a key candidate gene for the pork quality,which will provide theoretical basis for further studying the mechanism of fat deposition.
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