Expression and Hydroxylamine Cleavage of Thymosin Alpha 1 Concatemer

Abstract

Human thymosin alpha 1 (Tα1) is an important peptide in the development and senescence of immunological competence in human, and many studies have reported the expression of this peptide. In this study, we designed and synthesized theTα1gene according to theE. colicodon usage preference and constructed a 6×Tα1 concatemer. The latter was inserted into anE. coliexpression vector pET-22b (+), and transformed intoE. coliBL21 (DE3). After induction with IPTG, the concatemer protein was successfully expressed inE. colithen cleaved by hydroxylamine to release the Tα1 monomer. Gly-SDS-PAGE and mass spectrometry confirmed that the recombinant protein was cleaved as intended. The bioactivity of the Tα1 monomer was analyzed by lymphocyte proliferation and by mitochondrial activity in two different tumor cell lines. This study provides a description of the preparation of a bioactive Tα1, which may prove useful in future biomedical research.