Multi-Compartment and Multi-Host Vector Suite for Recombinant Protein Expression and Purification

Frontiers in Microbiology
Claudia OrtegaAgustín Correa

Abstract

Recombinant protein expression has become an invaluable tool in basic and applied research. The accumulated knowledge in this field allowed the expression of thousands of protein targets in a soluble, pure, and homogeneous state, essential for biochemical and structural analyses. A lot of progress has been achieved in the last decades, where challenging proteins were expressed in a soluble manner after evaluating different parameters such as host, strain, and fusion partner or promoter strength, among others. In this regard, we have previously developed a vector suite that allows the evaluation of different promoters and solubility enhancer-proteins, through an easy and efficient cloning strategy. Nonetheless, the proper expression of many targets remains elusive, requiring, for example, the addition of complex post-translation modifications and/or passage through specialized compartments. In order to overcome the limitations found when working with a single subcellular localization and a single host type, we herein expanded our previously developed vector suite to include the evaluation of recombinant protein expression in different cell compartments and cell hosts. In addition, these vectors also allow the assessment of alter...Continue Reading

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Citations

Jun 21, 2019·Protein Science : a Publication of the Protein Society·Germán L RosanoEduardo A Ceccarelli

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Methods Mentioned

BETA
glycosylation
PCR
Transfection
confocal microscopy
size-exclusion chromatography
gel filtration
size exclusion chromatography
by gel filtration
affinity purification

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