CLONING AND EXPRESSION OF PLANT LEGHEMOGLOBIN CDNA OF LUPINUS-LUTEUS IN ESCHERICHIA-COLI AND PURIFICATION OF THE RECOMBINANT PROTEIN

The yellow lupin leghemoglobin I gene (Ib1) was cloned in the pET-3a v ector. It was overexpressed in Escherichia coil BL21(DE3)pLysS cells, under the control of T7 RNA polymerase promoter. The recombinant LbI p rotein, containing E. coil derived heme, was purified to homogeneity u sing ion exchange and gel filtration chromatography. The recombinant L bI protein has spectral and immunochemical properties identical to LbI isolated from lupin root nodules. The identity between expressed poly peptide and native LbI protein was also supported by microsequencing a nalysis of the N-terminus of the purified recombinant protein.