A novel expression system for intracellular production and purification of recombinant affinity-tagged proteins in Aspergillus niger |
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Authors: | Andreas H F J Roth Petra Dersch |
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Institution: | (1) Institute of Microbiology, Technical University Braunschweig, Spielmannstrasse 7, 38106 Braunschweig, Germany;(2) Department of Molecular Infection Biology, Helmholtz Centre for Infection Research, Inhoffenstrasse. 7, 38124 Braunschweig, Germany; |
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Abstract: | A set of different integrative expression vectors for the intracellular production of recombinant proteins with or without
affinity tag in Aspergillus niger was developed. Target genes can be expressed under the control of the highly efficient, constitutive pkiA promoter or the novel sucrose-inducible promoter of the β-fructofuranosidase (sucA) gene of A. niger in the presence or absence of alternative carbon sources. All expression plasmids contain an identical multiple cloning sequence
that allows parallel construction of N- or C-terminally His6- and StrepII-tagged versions of the target proteins. Production of two heterologous model proteins, the green fluorescence
protein and the Thermobifida fusca hydrolase, proved the functionality of the vector system. Efficient production and easy detection of the target proteins
as well as their fast purification by a one-step affinity chromatography, using the His6- or StrepII-tag sequence, was demonstrated. |
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