pSS1-EBA140RIII–V

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	All constructs were verified by sequencing and subsequently transformed into Lactococcus lactis MG1363 by electroporation as described [22]. Five-ten colonies from each transformation were grown overnight at 30 °C in 5 ml LAB medium supplemented with 4% glycerol–phosphate, 5% glucose and 5 μg/ml erythromycin.

Protocol tips
All constructs were verified by sequencing and subsequently transformed into Lactococcus lactis MG1363 by electroporation as described [22]. Five-ten colonies from each transformation were grown overnight at 30 °C in 5 ml LAB medium supplemented with 4% glycerol–phosphate, 5% glucose and 5 μg/ml erythromycin.

Publication protocol

"All constructs were verified by sequencing and subsequently transformed into Lactococcus lactis MG1363 by electroporation as described [22].
Five-ten colonies from each transformation were grown overnight at 30 °C in 5 ml LAB medium supplemented with 4% glycerol–phosphate, 5% glucose and 5 μg/ml erythromycin. Culture supernatants were clarified by centrifugation at 9000g for 20 min and protein expression levels were assessed in the culture supernatants by ELISA using HRP-conjugated anti-His antibody (MACS, Miltenyi biotech, Germany) and by SDS-PAGE. L. lactis MG1363 harboring expression constructs was grown in a 1 l stirred bioreactors for 15 h at 30 °C [23]. Cells were removed by centrifugation at 9000 rpm for 30 min and the raw culture supernatant was clarified by filtration. Culture-filtrates were either used immediately or stored at − 20 °C."

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