Visfatin is a protein that is preferentially produced in visceral adipose tissue. Both its tissue expression and secreted plasma levels increase in parallel with obesity. Therefore, the present study is focused on detecting insertion in bovine visfatin gene and analyzing its effect on growth traits in four Chinese cattle breeds (Nangyang, Qinchuan, Jiaxian, and Chinese Holstein) using PCR-SSCP, DNA sequencing and agarose electrophoresis methods. For the first time, a 35-bp insertion was described in the gene and two alleles were revealed: Wand I. The chi2-test analysis demonstrated that all breeds were in agreement with Hardy-Weinberg equilibrium (P > 0.05). The associations of the novel 35bp-insertion of visfatin gene with growth traits of Nanyang cattle at 6, 12, 18, and 24 months old were analyzed: 6-month-old cows with genotype WW had greater birth weight than cows with genotype II and WI (P < 0.01 or P < 0.05). These results suggest that the presence of 35bp-insertion in visfatin gene may influence the birth weight and bodyweight in 6-month-old cows.