Construction of GST-β-catenin fusion protein expression vectors and the expression in prokaryotic cell

Objective To construct GST-β-catenin fusion protein expression vector and induce its expression in Escherichia coli(E.coli). Methods Total m RNA was extracted from COS-7 cells, and c DNA was formed by reverse transcription.Theβ-catenin coding sequence was amplified by polymerase chain reaction(PCR) and subcloned into p GEX-4T-2 vector. The positive recombinant was identified by restriction enzyme digestion and DNA sequencing. Then they were transformed into E.coli BL21, induced by IPTG and identified by SDS-PAGE and Western blot. Results The prokaryotic expression plasmid p GEX-4T-2-β-catenin was successfully constructed and confirmed by enzyme digestion and sequencing. The GST- β-catenin fusion proteins were expressed and confirmed by Western blot. Conclusion The prokaryotic expression plasmid ofβ-catenin was successfully constructed and the expression of fusion proteins in E.coli was confirmed. This study provides the basis for the further research on purifying Vinexin protein and the biological function ofβ-catenin.