Recombinant Rat Glucose-6-phosphate isomerase (Gpi)

In order to obtain a recombinant rat glucose-6-phosphate isomerase (Gpi) protein, the full-length mature rat Gpi encoding gene fragment (amino acids 2-558) along with an N-terminal 10xHis-tag and C-terminal Myc-tag sequence is inserted into an appropriate expression vector. The constructed vector is then introduced into E. coli cells through transformation. Following the selection of the transformed cells, a large-scale culture is initiated to allow for optimal protein expression. The N-terminal 10xHis-tag and C-terminal Myc-tag facilitate the subsequent purification and detection of the recombinant Gpi protein. Once the cells are lysed, the recombinant Gpi protein is extracted and subjected to purification. The purity of the recombinant Gpi protein is assessed using SDS-PAGE analysis, confirming its purity level of up to 90%. On the gel, the recombinant protein is observed as a distinct band with an approximate molecular weight of 66 kDa.