RESUMO
Thymosin ß4 (Tß4) is one of the most promising thymosins for future clinical applications, and it is anticipated that commercial demand for Tß4 will increase. In order to develop a new approach to produce recombinant Tß4, a 168 bp DNA (termed Tß4) was designed based on the Tß4 protein sequence and used to express a 4 × Tß 4 concatemer (four tandem copies of Tß4, termed 4 × Tß4) together with a histidine tag (6 × His) in E. coli (strain BL21). SDS-PAGE and western blot analysis were used to confirm that a recombinant 4 × Tß4 protein of the expected size (30.87 kDa) was produced following the induction of the bacterial cultures with isopropyl ß-D-thiogalactoside (IPTG). The E. coli-derived 4 × Tß4 was purified by Ni-NTA resin, and its activities were examined with regard to both stimulating proliferation of the mice spleen cells in vitro and in vivo wound healing. The results demonstrate that these activities of the E. coli-derived recombinant 4 × Tß4 were similar or even better than existing commercially obtained Tß4. This production strategy therefore represents a potentially valuable approach for future commercial production of recombinant Tß4.
