Expression and purification of recombinant T4 DNA ligase in E. coli

Abstract

In this study, we report results on the expression and purification of recombinant T4 DNA ligase. Plasmid pET16b-T4Dnl contains the gp30 gene which encodes for T4 DNA ligase. The target protein is fused with 10xHis tag to facilitate the purification and recovery. pET16b-T4Dnl was transformed into E. coli BL21(DE3) and then induced the expression of 10xHis-T4Dnl by IPTG. The recombinant protein was purified by Ni-NTA chromatography and confirmed by SDS-PAGE and Western blot. The activity of purified protein was tested by joining DNA λ/HindIII.