Fig. 1.

a Electrophoretogram showing PCR amplification of pyruvate kinase from the chromosomal DNA of S. aureus ATCC 12600. Lane M super mix marker obtained from Merek Biosciences Pvt Ltd. L1 Amplified 1.7 kb PCR product. b SDS-PAGE analysis of analysis pyruvate kinase: electrophoretogram showing the pyruvate kinase protein obtained from recombinant clone PK 1. Lane M High-molecular-weight marker from Merek Biosciences Pvt Ltd. Lane 1 Cytosolic fraction of clone PK 1. Lane 2 Cytosolic fraction of clone PK 1 induced with IPTG. Lane 3 Purified PK obtained by passing the cytosolic fraction of IPTG-induced PK 1 clone through Nickel metal chelate agarose column. c In vitro phosphorylation assay: SDS-PAGE gel was first stained with reagent A, followed by Coomasie brilliant blue R250 staining. Lane M High-molecular-weight marker from Merek Biosciences Pvt Ltd. Lane L1 phosphorylated PK obtained from Sephadex G-25 column. Lane L2 Pure PK obtained from Nickel metal chelate agarose column. d Western blot using an anti-His tag antibody: Lane L1 phosphorylated PK obtained from Sephadex G-25 column. Lane L2 Pure PK obtained from Nickel metal chelate agarose column