Standardization of qRT-PCR in analysis of gene expression of TLR receptors in swine
2011
Chytilová, M., University of Veterinary Medicine and Pharmacy in Košice (Slovak Republic) | Tkáčiková, Ľ. | Reichel, P.
The aim of this study was to optimise qRT -PCR that was used to analyse the influence of beta-glucans originating from Pleurotus ostreatus(pleuran, Natures) and Saccharomyces cerevisiae (Sigma) in three different concentrations (1 micro g per ml, 5 micro g per ml, 10 micro g per ml) on the changes of the gene expression of receptors TLR4 and TLR6 in the porcine intestinal epithelial line (IPEC -J2). Lipopolysaccharide (LPS) from E. coli(10 micro g per ml) was used as a control. The standardization of qRT -PCR included the selection of suitable primers and their optimal concentrations. The results of qRT -PCR revealed that lower concentrations of the tested glucans (1 micro g per ml and 5 micro g per ml) ailed to significantly affect the gene expression of the investigated receptors in IPEC -J2 cells. On the other hand, the stimulation of IPEC -J2 cells with pleuran at the highest concentration (10 micro g per ml), as well as stimulation of cells with LPS,doubled the expression of mRN A receptors TLR4 and TLR6.
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