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Figure 2 | Microbial Cell Factories

Figure 2

From: Dissection of an old protein reveals a novel application: domain D of Staphylococcus aureus Protein A (sSpAD) as a secretion - tag

Figure 2

Secretion competence of an sSpAD fusion protein. A: The sSpAD tag reacts with most secondary antibodies. Therefore sSpAD fusion proteins were detected on the same membrane as GroEL immuno blots. For the sSpAD immuno blots a secondary antibody was used which reacts against sSpAD. B: Anti GroEL immuno blots of the isolated fractions were performed to confirm that the periplasmic fractions obtained via osmolysis, were not contaminated with cytoplasmic proteins. C: Anti MalE immuno blots were performed to confirm that the isolated fractions correspond with the periplasm as the Maltose Binding Protein E is a periplasmic protein. -I: whole cell sample without induction of recombinant protein expression, +I: whole cell samples with induction of recombinant protein expression, P periplasmic fraction, C: cytoplasmic fraction; CCCP: carbonyl cyanide m-chlorophenylhydrazone, BL21(DE3): host strain, DADE: ΔtatABCDΔtatE host strain derived from MC4100.

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