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MOESM5 of A particular silent codon exchange in a recombinant gene greatly influences host cell metabolic activity

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posted on 2015-10-05, 05:00 authored by Natalie Rahmen, Christian Schlupp, Hitoshi Mitsunaga, Alexander Fulton, Tita Aryani, Lara Esch, Ulrich Schaffrath, Eiichiro Fukuzaki, Karl-Erich Jaeger, Jochen Büchs
Additional file 5. Product formation and sum of relative transcript abundances of four E. coli BL21(DE3) clones grown under inducing conditions. Soluble product formation (determined using a BioLector device) and sum of relative transcript abundances (determined from conventional shake flasks) as function of time for four E. coli BL21(DE3) clones belonging to respiration behavior Type A (Leu143-CTT, -TTG; white background) and respiration behavior Type B (Leu143-CTA, -CTG; grey background) during cultivation in Wilms-MOPS mineral autoinduction medium containing 0.5 g/L glucose, 5 g/L glycerol, 2 g/L lactose. ‘lipA vs. cysG’ represents the sum of the relative transcript abundance of the plasmid-encoded target gene lipA (encoding Bacillus subtilis lipase A) in relation to the genome-encoded reference gene cysG (encoding uroporphyrin III C-methyltransferase). ‘bla vs. cysG’ represents the relative plasmid copy number as the sum of the relative transcript abundance plasmid-encoded reference gene bla (encoding beta-lactamase) in relation to the genome-encoded reference gene cysG. ‘lipA vs. bla’ represents the factor of lipA induction as transcript abundance of the plasmid-encoded target gene lipA in relation to the plasmid-encoded reference gene bla. Cultivation conditions: 37 °C, 250 mL flasks, filling volume 10 mL, shaking frequency 350 rpm, shaking diameter 50 mm (in conventional flasks); 37 °C, 48-well Flowerplate, filling volume 1 mL, shaking frequency 1000 rpm, shaking diameter 3 mm (in BioLector).

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