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The Escherichia coli-derived thymosin ?4 concatemer promotes cell proliferation and healing wound in mice.


ABSTRACT: Thymosin ?4 (T?4) is one of the most promising thymosins for future clinical applications, and it is anticipated that commercial demand for T?4 will increase. In order to develop a new approach to produce recombinant T?4, a 168 bp DNA (termed T?4) was designed based on the T?4 protein sequence and used to express a 4 × T? 4 concatemer (four tandem copies of T?4, termed 4 × T?4) 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 × T?4 protein of the expected size (30.87 kDa) was produced following the induction of the bacterial cultures with isopropyl ?-D-thiogalactoside (IPTG). The E. coli-derived 4 × T?4 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 × T?4 were similar or even better than existing commercially obtained T?4. This production strategy therefore represents a potentially valuable approach for future commercial production of recombinant T?4.

SUBMITTER: Wang X 

PROVIDER: S-EPMC3671520 | BioStudies | 2013-01-01

REPOSITORIES: biostudies

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