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Suppl. F igure 2

Suppl. F igure 2. Vector of expression. Cytoplasmic : pSNAP , pSORTc , pFc. P eriplasmic : pHEN2, pSORTp. Transformation in BL21 + Sulfhydryl oxydase LB + antibiotics ( ABs ). Transformation in TG1 2 xTY +1%glucose+ AB. No colonies. No colonies. Colonies. C olonies.

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Suppl. F igure 2

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  1. Suppl. Figure 2 Vector of expression Cytoplasmic : pSNAP,pSORTc, pFc Periplasmic : pHEN2, pSORTp Transformation in BL21 + Sulfhydryl oxydase LB +antibiotics (ABs) Transformation in TG1 2 xTY +1%glucose+ AB No colonies No colonies Colonies Colonies Sample 2-3 colonies in 3 mL LB, grow ON at 30°C, 210 rpm Check ABs Competent cells Check ABs Competent cells Makeglycerol Stocks Prepare 5 mL ON culture LB + AB Prepare 30 mL ON culture 2xTY+ 1%glu+ AB Growat 30°C Growat 37°C Add 2mL pre-culture in 500 mL LB+ ABs Add 25mL pre-culture in 500 mL 2xTY+ 0.1%glu+ AB Growuntil OD600nm=0.4 Growuntil OD600nm=0.6 Add arabinose 0.5% w/v, decrease the temperature to 20°C Add IPTG 0.5mM Growuntil OD600nm=0.6 Growat 20°C Add IPTG 0.05mM Grow 4 h at 28°C Harvestbacteria by centrifugation at 4000 rpm, 15 min, 4°C freeze and store at -20°C Grow ON Harvestbacteria by centrifugation at 4000 rpm, 15 min, 4°C ,freeze and store at -20°C

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