scholarly journals PROFICS: A bacterial selection system for directed evolution of proteases

2021 ◽  
pp. 101095
Author(s):  
Christina Kröß ◽  
Petra Engele ◽  
Bernhard Sprenger ◽  
Andreas Fischer ◽  
Nico Lingg ◽  
...  
Keyword(s):  
Directed Evolution ◽  
Selection System ◽  
Bacterial Selection

A bacterial selection for the directed evolution of pyruvate aldolases

2004 ◽  
Vol 12 (15) ◽  
pp. 4067-4074 ◽  
Author(s):  
Jennifer S Griffiths ◽  
Manoj Cheriyan ◽  
Jayme B Corbell ◽  
Luka Pocivavsek ◽  
Carol A Fierke ◽  
...  
Keyword(s):  
Directed Evolution ◽  
Selection For ◽  
Bacterial Selection

scholarly journals An in vivo selection system with tightly regulated gene expression enables directed evolution of highly efficient enzymes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Parinthon Nearmnala ◽  
Manutsawee Thanaburakorn ◽  
Watanalai Panbangred ◽  
Pimchai Chaiyen ◽  
Narupat Hongdilokkul
Keyword(s):  
Directed Evolution ◽  
Selection Pressure ◽  
Catalytic Efficiency ◽  
Selection System ◽  
In Vivo Selection ◽  
Highly Efficient ◽  
Highly Active ◽  
Wide Range ◽  
Regulated Gene Expression

AbstractIn vivo selection systems are powerful tools for directed evolution of enzymes. The selection pressure of the systems can be tuned by regulating the expression levels of the catalysts. In this work, we engineered a selection system for laboratory evolution of highly active enzymes by incorporating a translationally suppressing cis repressor as well as an inducible promoter to impart stringent and tunable selection pressure. We demonstrated the utility of our selection system by performing directed evolution experiments using TEM β-lactamase as the model enzyme. Five evolutionary rounds afforded a highly active variant exhibiting 440-fold improvement in catalytic efficiency. We also showed that, without the cis repressor, the selection system cannot provide sufficient selection pressure required for evolving highly efficient TEM β-lactamase. The selection system should be applicable for the exploration of catalytic perfection of a wide range of enzymes.

scholarly journals Directed evolution of a transcription factor PbrR to improve lead selectivity and reduce zinc interference through dual selection

2020 ◽  
Author(s):  
Xiaoqiang Jia ◽  
Yubing Ma ◽  
Rongrong Bu ◽  
Tingting Zhao ◽  
Kang Wu
Keyword(s):  
Transcription Factor ◽  
Directed Evolution ◽  
Metal Binding ◽  
Binding Specificity ◽  
Selection Marker ◽  
Selection System ◽  
Lead And Cadmium ◽  
Negative Selection Marker ◽  
Binding Residue ◽  
Ampicillin Resistance Gene

Abstract Directed evolution has been proven as a powerful tool for developing proteins and strains with novel or enhanced features. In this study, a dual selection system was designed to tune the binding specificity of a transcription factor to a particular ligand with the ampicillin resistance gene amp (ON selection) as the positive selection marker and the levansucrase gene sacB (OFF selection) as the negative selection marker. It was applied to the lead responsive transcription factor PbrR in a whole-cell lead biosensor previously constructed in our lab. After multiple rounds of ON-OFF selection, two mutants with higher specificity for lead were selected. Structural analysis revealed that the mutation C134 located on the metal-binding loop at the C-terminal of PbrR is likely associated with the enhanced binding to both lead and cadmium. The double mutations D64A and L68S close to the metal-binding residue C79 may lead to the reduced binding specificity toward zinc ions. This dual selection system can be applied to engineer the specificity of other transcription factors and provide fine-tuned tools to synthetic biology.

scholarly journals A generic selection system for improved expression and thermostability of G protein-coupled receptors by directed evolution

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Christoph Klenk ◽  
Janosch Ehrenmann ◽  
Marco Schütz ◽  
Andreas Plückthun
Keyword(s):  
Directed Evolution ◽  
G Protein ◽  
G Protein Coupled Receptors ◽  
Selection System ◽  
G Protein Coupled

Switching the Ligand Specificity of the Biosensor XylS from meta to para-Toluic Acid through Directed Evolution Exploiting a Dual Selection System

2019 ◽  
Vol 8 (12) ◽  
pp. 2679-2689
Author(s):  
Yuki Ogawa ◽  
Yohei Katsuyama ◽  
Kento Ueno ◽  
Yasuo Ohnishi
Keyword(s):  
Directed Evolution ◽  
Ligand Specificity ◽  
Selection System

Pilot selection system development.

1978 ◽  
Author(s):  
David R. Hunter ◽  
Nancy A. Thompson
Keyword(s):  
System Development ◽  
Selection System ◽  
Pilot Selection

Two validation studies of a school teacher selection system

2008 ◽  
Author(s):  
Drozd Mutual ◽  
Paul Kirk ◽  
Theodore Hayes
Keyword(s):  
Validation Studies ◽  
Teacher Selection ◽  
School Teacher ◽  
Selection System

Validation of a School Teacher Selection System

2006 ◽  
Author(s):  
Dee W. Drozd ◽  
Theodore L. Hayes
Keyword(s):  
Teacher Selection ◽  
School Teacher ◽  
Selection System
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