Cloning, expression, purification and biologic characteristics of Tα1 ② protein
AIM: To clone, express and purify recombinant Thymosin alpha 1 ② protein (Tα1②) in E. coli. METHODS: A 189 bp recombinant gene of 2 × Tα1 tandem repeats was synthesized and cloned into pET-22b( + ) vector. The expression plasmid was then transformed into E. coll. BL21 ( DE3 ). The expression of the recombinant protein Tα1② was induced with IPTG and detected by SDS-PACE and Western blot. The biological activity of Tα1② was identified by MTT assay. RESULTS: A novel protein with expected molecular mass was expressed. The expressed product could be recognized by a monoclonal antibody against the human Tα1. The purified Tα1② stimulated the proliferation of mice splenic lymphocytes. CONCLUSION: The recombinant Tα1② is anccessfully cloned, expressed and purified. The purified Tα1 ②protein has the same function as synthesized Tα1 but higher bioactivity.
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维普
