Scaling production of GFP1-10 detector protein in E. coli for secretion screening by split GFP assay.

ABSTRACT:

Background

The split GFP assay is a well-known technology for activity-independent screening of target proteins. A superfolder GFP is split into two non-fluorescent parts, GFP11 which is fused to the target protein and GFP1-10. In the presence of both, GFP1-10 and the GFP11-tag are self-assembled and a functional chromophore is formed. However, it relies on the availability and quality of GFP1-10 detector protein to develop fluorescence by assembly with the GFP11-tag connected to the target protein. GFP1-10 detector protein is often produced in small scale shake flask cultivation and purified from inclusion bodies.

Results

The production of GFP1-10 in inclusion bodies and purification was comprehensively studied based on Escherichia coli as host. Cultivation in complex and d

SUBMITTER: Muller C 

PROVIDER: S-EPMC8482599 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature