The Study On The Expression Of Monosaccharide Transporters GLUT2Regulated By Transcription Factors GATA3and USF-1

Monosaccharide transporter GLUT2is one of the important carriers for the delivery of glucose and fructose in small intestine. Its expression directly affect the absorption of monosaccharide nutrients in intestine and ultimately affect the growth rate and feed conversion rate (FCR). Our research focuses on the expression and regulation mechanism of the transcription factor GATA3and USF-1, which binding5’upstream of the GLUT2. The information of chicken GATA3and USF-1were analysis to provide the reference for the structure and function of the protein. The GATA3and USF-1gene were transfected into293T cells by pcDNA3.1vector. The experimental set mock-vehicle group, transfected group and untransfected group. Observed the morphological changes after transfection48h and extracted total RNA from cells. The absolute expression of target gene and GLUT2in the transfected cells were tested by Real-time PCR.Results:the transcription factor GATA3consists of444amino acids and USF-1consists of310amino acids through bioinformatics software ExPASy analysis. Both are hydrophilic nuclear proteins, unstable, without signal peptide. Through analysis of online software GOR4to obtain the secondary structure model of GATA3and USF-1. Successfully constructed the eukaryotic expression vector pcDNA3.1-GATA3and pcDNA3.1-USF-1and expressed in293T cells. In the experiment of pcDNA3.1-GATA3transfected293T cells, transfected group compared with mock-vehicle groups and untransfected group, the expression of GATA3is very significant difference (P<0.01), while the expression of GLUT2in three groups were not significant (P>0.05); In the experiment of pcDNA3.1-USF-1transfected293T cells, transfected group compared with mock-vehicle groups and untransfected group demonstrate the expression of GATA3is significant difference (0.01<P<0.05), while the expression of GLUT2in three groups were not significant (P>0.05). After statistical analysis, we draw the following conclusion:transcription factors GATA3and USF-1expressed in239T cells failed to increase the expression of monosaccharide transporter GLUT2. This study establishes a theoretical and practical foundation for the further study on GATA3and USF-1regulated the expression of GLUT2and elucidates the mechanism of nutrient absorption in chicken intestine.