To know the relation on concentrate to forage ratio of pellet feed and gene expression of GLUT1, FATP4,PepT1 mRNA in different tissues. 12 Holstein bull calves with the similar age and weight were chosed and divided into 4 groups with 3 calves. According to NRC(2001), calves in each group were fed the pellet feed with the concentrate to forage with ratio of 75∶25(I), 70∶30(Ⅱ), 65∶35(Ⅲ) and 60∶40(Ⅳ) which were made from the quality ratio of alfalfa to triticale for 3∶1 by 20 type granulated feed unit. The experimental time included 14 days of preliminary trial period and 56 days of the experimental period. The results showed that 1) The expression of GLUT1 mRNA in liver, rumen, duodenum and cecum in group Ⅲ were significantly higher than that of group Ⅳ (P<0.05).The expression of GLUT1 mRNA in abomasum of group I were significantly higher than that of other groups (P<0.05). 2) The expression of FATP4 mRNA in liver, cecum of group Ⅲ and rumen, duodenum, abomasum of group I were higher than that of other groups. 3) The expression of PepT1 mRNA in rumen, duodenum, abomasum of group I , liver of group Ⅳ and cecum of group Ⅲ were higher than that of other groups. In conclusion, the different concentrate to forage ratio in pellet feed could influence the expression of GLUT1, FATP4 and PepT1 in different tissues. The concentrate to forage ratio with 65∶35 was beneficial for calves to absorb the glucose, fat acid and protein.