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Figure 4 | Virology Journal

Figure 4

From: Expressing gK gene of duck enteritis virus guided by bioinformatics and its applied prospect in diagnosis

Figure 4

The different samples were analyzed by Coomassie blue-stained polyacrylamide gel. The direction of arrow represented the target protein. M represented standard protein molecular weight markers. A. Lane 1, the un-induced BL21 bacteria within pET-32b(+) plasmid; Lane 2, the induced BL21 bacteria within pET-32b(+) plasmid; Lane 3, the un-induced BL21 bacteria within pET-32b(+)/fgK plasmid; Lane 4, the supernatant of IB with sonication was analyzed by SDS-PAGE, that the IB was harvested from the induced BL21 bacteria within pET-32b(+)/fgK plasmid; Lane 5, the pellet of IB with sonication was analyzed by SDS-PAGE, that the IB was harvested from the induced BL21 bacteria within pET-32b(+)/fgK plasmid; Lane 6, the induced Plys bacteria within pET-32b(+)/fgK plasmid; Lane 7, the induced Rosetta bacteria within pET-32b(+)/fgK plasmid. B. Lane 1, the un-induced BL21 bacteria within pET-32b(+) plasmid; Lane 2, the induced BL21 bacteria within pET-32b(+) plasmid; Lane 3, the un-induced BL21 bacteria within pET-32b(+)/tgK plasmid; Lane 4, the induced BL21 bacteria within pET-32b(+)/tgK plasmid; Lane 5, the induced Plys bacteria within pET-32b(+)/tgK plasmid; Lane 6, the induced Rosetta bacteria within pET-32b(+)/tgK plasmid. C. The determination of the optimal induced temperature. Lanes 1, 2 and 3 representing the induced temperatures were 25°C, 30°C and 37°C, respectively. D. The determination of the optimal induced IPTG final concentration. Lanes 1, 2, 3, 4, 5, 6, 7 and 8 representing the induced IPTG final concentrations were 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM and 1.0 mM, respectively. E. The determination of the optimal induced duration. Lanes 1, 2, 3 and 4 representing the induced durations were 2.0 h, 4.0 h, 8.0 h and 16.0 h, respectively. F. Purification of IB and recombinant truncated gK. Lanes 1 and 2 respectively represented the supernatant and pellet of the induced bacteria within pET-32b(+)/tgK plasmid with sonication; Lane 3 represented the purified recombinant truncated gK by a single chromatographic step of IMAC on Ni2+-NTA agarose.

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