Recombinant Mycobacterium tuberculosis Amidophosphoribosyltransferase (purF)

To generate a recombinant Mycobacterium tuberculosis Amidophosphoribosyltransferase (purF) protein in the E. coli expression system, the full-length mature purF gene fragment (35-527aa) specific to Mycobacterium tuberculosis along with an N-terminal 6xHis-tag is cloned into a suitable expression vector. The resulting recombinant plasmid is then transformed into E. coli host cells using a transformation method. The transformed cells are selected using antibiotic resistance markers to ensure the presence of the recombinant plasmid and are cultured in a growth medium optimized for protein expression. The cells are induced with an inducer molecule, such as IPTG, to initiate the expression of the recombinant purF protein. After incubation, the cells are harvested and lysed to release the intracellular contents including the expressed recombinant purF protein. The purity of the recombinant purF protein is assessed through SDS-PAGE analysis, demonstrating a purity level of up to 85%. On the gel, the recombinant Mycobacterium tuberculosis purF protein appears as a distinct band with a molecular weight of approximately 57 kDa.