Kód: 06825626
A recombinant S100A4 protein was produced in Pichia §pastoris (Pichia). The expression vector contained §parts of the AOX1 promoter and also a secretion §signal, making it possible to purify the protein §from the yeast media after ... celý popis
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A recombinant S100A4 protein was produced in Pichia §pastoris (Pichia). The expression vector contained §parts of the AOX1 promoter and also a secretion §signal, making it possible to purify the protein §from the yeast media after methanol§induction. Attempts were made to purify the protein §from the media both by hydrophobic interaction§chromatography (HIC) and by anionic ion exchange §chromatography (IEC). The binding capacity to§the hydrophobic material was rather low, but the §eluted protein did however show an acceptable purity§for utilization in downstream functional assays. The §protein eluted from the hydrophobic column was §labelled with the fluorescent dye (Cy3) and used for §internalization studies in the human cell line HeLa. §Similar to a mouse rS100A4 produced in bacterial §cells, the Pichia produced protein was also found §internalized into the HeLa cells. The biological §significance of these findings is at present not §elucidated and will be further§studied.
Zařazení knihy Knihy v angličtině Mathematics & science Biology, life sciences Biochemistry
1648 Kč
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