Thoroughbred is a breed of horse mainly used in racing horse industry. Thoroughbred is also used as an important model in the field of exercise and physiology because of their physical characteristics such as tenacity and speediness. Although many studies were focused on physical and physiological adaptations, the regulatory pathways and mechanisms of targeted genes are still remained to be uncovered in race horse. The purpose of this study is to analyze the molecular and phylogenetic characteristics of the Krüppel-like factor (KLF) family genes of Equus caballus. As results, Zinc finger domain and coding sequence (CDS) of Thoroughbred KLF family protein are highly conserved among species. Also, quantitative real time polymerase chain reaction (qRT-PCR)　was conducted to investigate the mRNA expression levels of horse KLF3, 5, 7, 10, 12 and 16 in different horse tissues. As a result, six KLF genes showed high expression in the cecum and high expression in lung tissue, indicating tissue-specific expression. In conclusion, these results indicate that horse KLF family genes are highly conserved during evolution and will be used as a valuable information for further study of horse KLF genes.