According to the transcriptome data of Gossypium barbadense, we cloned GbTCP5 gene from cotton (Gossypim barbadense L.) cultivar ‘Xinhai 21’using reverse transcription PCR methods. GbTCP5 gene contains open reading frame (ORF) of 945 bp in length, 314 amino acids residues with a predicted molecular mass of about 34.95 kD and a basic isoelectric point of 8.41 with a highly conservation TCP domain in the encoded putative protein. Amino acid sequence alignment revealed of GbTCP5 proteins shared high degree of identity with other higher plant TCP proteins and the gene in evolution has proved to be conservative. Subcellular localization showed that GbTCP5 expressed possibly in nucleus. The phylogenetic tree showed that GbTCP5 was located at the same distribute with GhTCP17. The result of real time PCR exhibited that GbTCP5 had a higher expression level in fibers(35d), may be involved in secondary wall synthesis. GbTCP5 was grown on SDTrpHisAde deficient medium and the Xgal test was blue. The results showed that GbTCP5 had the function of transcriptional activation in yeast.