Taiping chicken is indigenous chickens (Gallus gallus domesticus), which was one of China's excellent poultry species, is an excellent chicken in Gansu Province. As the problems caused by the overuse of antibiotics become more and more severe, people begin to look for ways to replace them. Among them, probiotics and fructo-oligosaccharides are the research hotspot to replace antibiotics. Probiotics and fructo-oligosaccharides can promote the absorption of nutrients, improve the ability to resist and prevent diseases, and improve the intestinal tissue morphology. In this study, we used RNA-Seq analysis to study the gene expression in ileum tissue after Taiping chicken was given probiotics and fructo-oligosaccharides. In total, 67 genes were differentially expressed in the ileum. Ten of the differently expressed genes were further validated by RT-qPCR. In addition, these differentially expressed genes were mainly enriched to tyrosine metabolism, AGE-RAGE signaling pathway in diabetic complications, phenylalanine metabolism, and pyrimidine metabolism. The results which this study provides contribute to our understanding application of probiotics and fructo-oligosaccharides in indigenous chickens production and provide a theoretical basis for the genetic development of indigenous chickens.