Molecular cloning, expression, purification and osteoblasts proliferation activity of sika deer thymosin beta10

Thymosin beta 10 (Tβ10) is a member of the β-thymosin family. As an actin-binding peptide, thymosin β10 is involved in many important biological activities. Transcriptome sequencing results suggest that Tβ10 may play important roles in the growth of deer antler. In this study, Tβ10 cDNA was isolated from sika deer, and complete open reading frame consisting of 129 nucleotides was obtained by PCR amplification. The predicted peptide was 42 amino acids in length. The sdTβ10 cDNA was cloned and expressed in Escherichia coli resulting in a 6 kDa recombinant-His tagged protein. The recombinant, non-glycosylated peptide was overexpressed in a soluble form and purified by immobilized metal ion affinity chromatography. Functional studies revealed that recombinant Tβ10 stimulated osteoblasts proliferation. This study provides the first evidence that recombinant sika deer Tβ10 promotes proliferation in an osteoblasts cell model.