Abstract: Three primers were designed to amplify the fragments of exon10 to exon14, exon13 to exon17 and exon16 to exon19 in alpacas. These fragments were 414, 539 and 426 bp, respectively. 1 044 bp cDNA sequence of KIT gene, spanned from exon10 to exon19, was obtained by contig software, and submitted to GenBank (DQ450844). The nucleotide sequence was aligned with the corresponding part of other animals, and the similarity was above 94%, which indicated that the sequences were homologous. It was confirmed that the sequence obtained in this study was the KIT gene of alpacas. And the deduced polypeptide was got by the translation software and it coded 347 amino acids. The multiple sequence alignments revealed that the similarity of alpacas KIT sequence were above 95% with other species. Three amino acid substitutions were detected in the polypeptide coded by exon1019 of KIT gene in alpacas, which were I→V at the position of 2, S→P at the position of 201 and K→R at the position of 344. And bioinformatics analysis indicated that there was much alpha-helix in the secondary structure; And the protein encoded the mast/stem cell growth factor receptor. These results would be the foundation of further researching the association between KIT gene and coat color of alpacas.