Construction of GST-LMO1 Fusion Protein Expression Vectors and the Expression in Prokaryotic Cell

Objective To construct GST-LMO1 fusion protein expression vector and induce its expression in Escherichia coli(E.coli).Methods The coding sequence of LMO1 and its deletion fragments were amplified from the human fetal brain as the template by PCR and inserted into pGEX-5X-2 by BamHⅠand EcoRⅠ.The positive recombinants were identified by restriction endonuclease 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 pGEX-5X-2-LMO1 and its deletion mutants were successfully constructed and confirmed by enzyme digestion and sequencing.The GST-LMO1 fusion proteins were expressed and confirmed by Western blot.Conclusion The prokaryotic expression plasmid of LMO1 and its deletion mutants were successfully constructed and the expression of fusion proteins in Escherichia coli was confirmed.This study provides the basis for the further research on the structure and function of LMO1.