Modular Protein Expression Toolbox (MoPET), a standardized assembly system for defined expression constructs and expression optimization libraries

PloS One
Ernst WeberMark Trautwein

Abstract

The design and generation of an optimal expression construct is the first and essential step in in the characterization of a protein of interest. Besides evaluation and optimization of process parameters (e.g. selection of the best expression host or cell line and optimal induction conditions and time points), the design of the expression construct itself has a major impact. However, the path to this final expression construct is often not straight forward and includes multiple learning cycles accompanied by design variations and retesting of construct variants, since multiple, functional DNA sequences of the expression vector backbone, either coding or non-coding, can have a major impact on expression yields. To streamline the generation of defined expression constructs of otherwise difficult to express proteins, the Modular Protein Expression Toolbox (MoPET) has been developed. This cloning platform allows highly efficient DNA assembly of pre-defined, standardized functional DNA modules with a minimal cloning burden. Combining these features with a standardized cloning strategy facilitates the identification of optimized DNA expression constructs in shorter time. The MoPET system currently consists of 53 defined DNA modules d...Continue Reading

References

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Citations

Dec 11, 2020·Metabolic Engineering·Rosanna YoungPaul S Freemont
Feb 3, 2021·Trends in Biotechnology·Roberto Di BlasiFrancesca Ceroni

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Methods Mentioned

BETA
PCR
transfection
ELISA

Software Mentioned

MoClo
GoldenBraid
Modular
MoPET

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