ObjectiveTo construct an engineering strain of pMAL- C2x - Tα1/TB1 which can express Thymosin α1(Tα1).MethodsTα1 gene was synthesized and amplified by PCR, PCR product and pMAL- C2x vector were digested with restriction endonuclease BamHI and EcoR I respectively, and were linked with T4 DNA ligase to construct pMAL- C2x - Tα1.After being sequenced, pMAL- C2x - Tα1 vector was transformed into E.coli TB1 for fusion expression under induction of IPTG.. The expressed product was identified by Western blot. ResultsThe DNA sequence of the synthesized Tα1 gene was identical to the original design.The constructed recombinant plasmid pMAL- C2x - Tα1 was highly expressed in E.coli TB1. The BMP - Tα1 fusion protein was about 33.6% of the total bacteria protein and the relative molecular weight of BMP - Tα1 fusion protein was about 45×10³.ConclusionThe successful construction and expression of pMAL- C2x - Tα1/TB1 laid a foundation for the research of purification and biological acitivity of recombinant Tα1.