Modern biotechnology has generated an impressive set of molecular tools: for instance the ability to generate large sets of error prone mutant libraries or cDNA libraries. When these libraries are expressed in a host (e.g. Aspergillus niger) not all strains produce a secreted protein. This is mainly dependent on the quality of the library. Here we describe a novel expression system that was developed by using genome expression profiling under different conditions. We were able to identify promoters that fit the required expression profile. These promoters were both up regulated during protein secretion and were not expressed during overexpression of intracellular proteins. By making use of transcriptomics for useful promoter identification, we were able to generate reporter construct(s) that allow us to easily select clones that secrete proteins. This technology can speed up novel protein discovery significantly. Additionally we have shown this approach is not limited to fungi but can also be applied to other production organisms.
Full conference title:
10th EUROPEAN CONFERENCE ON FUNGAL GENETICS
- ECFG 10th (2010)