A novel expression system for intracellular production and purification of recombinant affinity-tagged proteins in Aspergillus niger

作者：Roth Andreas H F J; Dersch Petra^*

来源：Applied Microbiology and Biotechnology, 2010, 86(2): 659-670.

DOI：10.1007/s00253-009-2252-9

• Aspergillus niger
• Affinity-tagged proteins
• Protein production
• Inducible promoter
• Expression vectors
• sucA promoter

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 beta-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 His(6)- 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 His(6)- or StrepII-tag sequence, was demonstrated.

• 出版日期2010-3