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Fig. 1 | Microbial Cell Factories

Fig. 1

From: PglB function and glycosylation efficiency is temperature dependent when the pgl locus is integrated in the Escherichia coli chromosome

Fig. 1

Generation of functional SDB1pgl integrants. A Schematic diagram depicting the pSECpgl suicide plasmid. B Strategy for integration of the C. jejuni pgl locus in the SDB1 genome via allelic exchange. C Confirmation of correct integration of the pgl locus via PCR in five SDB1pgl clones. Oligos 1 and 6 align outside of the chosen homology arms, while oligos 2 and 5 align to the pgl locus and the KanR tag, respectively. Formation of amplicons 1&2 and 5&6 is possible only in case of successful integration in the correct location. D SDB1pgl integrants can glycosylate a model carrier protein (G-NetB). SDB1 transformed with a plasmid encoding pgl locus (pACYCpgl) as well as a plasmid encoding pgl locus with a non-functional PglB (pACYCpgl::pglB) were used as positive and negative controls, respectively. His-pulldown followed by western blotting confirms glycoprotein production

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Microbial Cell Factories

ISSN: 1475-2859

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