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(IPTG(-/+) = without/with 0.2 mM IPTG for 3-6 hours, adding IPTG to a bacteria culture with OD600 0.2-0.3. M: Protein molecular weight marker ladder. Lane 2~10、13~15: pET28 plasmids encoding crtIEB separated by self-cleaving ribozyme, crtI+crtB+cr...) |
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Latest revision as of 14:52, 8 October 2022
IPTG(-/+) = without/with 0.2 mM IPTG for 3-6 hours, adding IPTG to a bacteria culture with OD600 0.2-0.3. M: Protein molecular weight marker ladder. Lane 2~10、13~15: pET28 plasmids encoding crtIEB separated by self-cleaving ribozyme, crtI+crtB+crtE, crtI&crtB&crtE,crtB+crtE+crtY,crtY,crtB without any tag were transformed into BL21(DE3) Rosetta strain, single clones(IEB4,IEB5,IEB6,I7,B1,E1,BEY2,Y1,B1)were picked for liquid LB culture. Lane 11~12: pET28 plasmids encoding crtBE separated by self-cleaving ribozyme without any tag were transformed into BL21(DE3) Hi-control Rosetta strain, single clones(BE12)were picked for liquid LB culture.
Protein expression was induced in parallel cultures by IPTG. Bacterial cultures were monitored by OD600, and 5x10^7 cells were harvested by centrifugation and lysis in 1x SDS sample buffer. Equal amount (10 μL, 2x10^6 cells) of whole cell lysate were analyzed by SDS-PAGE (14% separation gel).
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current | 14:52, 8 October 2022 | 1,702 × 1,116 (2.65 MB) | Atlantics (Talk | contribs) | ||
13:28, 6 October 2022 | 1,474 × 1,036 (1.83 MB) | Atlantics (Talk | contribs) | IPTG(-/+) = without/with 0.2 mM IPTG for 3-6 hours, adding IPTG to a bacteria culture with OD600 0.2-0.3. M: Protein molecular weight marker ladder. Lane 2~10、13~15: pET28 plasmids encoding crtIEB separated by self-cleaving ribozyme, crtI+crtB+cr... |
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