Enhanced and Secretory Expression of Human Granulocyte Colony Stimulating Factor by Bacillus subtilis SCK6

Figure 2

(a) Restriction analysis of pNWPH-GCSF expression plasmid resolved on 1% agarose gel. M, 1 kb DNA size marker; Lane 1, undigested pNWPH-GCSF; Lane 2, pNWPH-GCSF after double digestion with NdeI and HindIII restriction endonucleases. (b) 13% SDS-PAGE analysis of TCA-acetone precipitated culture supernatant of transformed B. subtilis SCK6. Lane M represents protein size marker; Lanes 1–7, sample fractions collected at 24, 36, 48, 60, 72, 84, and 96 hours of cell growth. (c) Growth of recombinant B. subtilis harboring pNWPH-GCSF in 2x L-Mal medium. -axis shows time in hours while primary -axis reflects the concentration of GCSF (μg/mL) in culture supernatant, and secondary -axis shows cell growth, monitored by absorbance measurement at 600 nm.

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