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Protein Engineering

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Cover of 'Protein Engineering'

Table of Contents

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    Book Overview
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    Chapter 1 Protein Engineering: Past, Present, and Future
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    Chapter 2 Rational and Semirational Protein Design
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    Chapter 3 Computational Analysis of Protein Tunnels and Channels
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    Chapter 4 YASARA: A Tool to Obtain Structural Guidance in Biocatalytic Investigations
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    Chapter 5 A Computational Library Design Protocol for Rapid Improvement of Protein Stability: FRESCO
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    Chapter 6 Directed Evolution of Proteins Based on Mutational Scanning
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    Chapter 7 A Brief Guide to the High-Throughput Expression of Directed Evolution Libraries
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    Chapter 8 Library Growth and Protein Expression: Optimal and Reproducible Microtiter Plate Expression of Recombinant Enzymes in E. coli Using MTP Shakers
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    Chapter 9 Normalized Screening of Protein Engineering Libraries by Split-GFP Crude Cell Extract Quantification
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    Chapter 10 Functional Analysis of Membrane Proteins Produced by Cell-Free Translation
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    Chapter 11 Practical Considerations Regarding the Choice of the Best High-Throughput Assay
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    Chapter 12 High-Throughput Screening Assays for Lipolytic Enzymes
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    Chapter 13 Continuous High-Throughput Colorimetric Assays for α -Transaminases
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    Chapter 14 Colorimetric High-Throughput Screening Assays for the Directed Evolution of Fungal Laccases
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    Chapter 15 Directed Coevolution of Two Cellulosic Enzymes in Escherichia coli Based on Their Synergistic Reactions
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    Chapter 16 Program-Guided Design of High-Throughput Enzyme Screening Experiments and Automated Data Analysis/Evaluation
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    Chapter 17 Solid-Phase Agar Plate Assay for Screening Amine Transaminases
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    Chapter 18 Ultrahigh-Throughput Screening of Single-Cell Lysates for Directed Evolution and Functional Metagenomics
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    Chapter 19 Isolation of pH-Sensitive Antibody Fragments by Fluorescence-Activated Cell Sorting and Yeast Surface Display
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    Chapter 20 Library Generation and Auxotrophic Selection Assays in Escherichia coli and Thermus thermophilus
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    Chapter 21 Erratum to: Functional Analysis of Membrane Proteins Produced by Cell-Free Translation
Attention for Chapter 9: Normalized Screening of Protein Engineering Libraries by Split-GFP Crude Cell Extract Quantification
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Chapter title
Normalized Screening of Protein Engineering Libraries by Split-GFP Crude Cell Extract Quantification
Chapter number 9
Book title
Protein Engineering
Published in
Methods in molecular biology, January 2018
DOI 10.1007/978-1-4939-7366-8_9
Pubmed ID
Book ISBNs
978-1-4939-7364-4, 978-1-4939-7366-8
Authors

Javier Santos-Aberturas, Mark Dörr, Uwe T. Bornscheuer, Santos-Aberturas, Javier, Dörr, Mark, Bornscheuer, Uwe T.

Abstract

The different expression level and solubility showed by each protein variant represents an important challenge during screening campaigns: Usually, the total activity measurement constitutes the only criterion for identifying improved variants. This hampers the chances of finding interesting mutants, especially if the aim is to improve activity: On the one hand, interesting but poorly soluble variants will remain undetectable. On the other hand, a mutation might not increase activity, but improve expression level or solubility. The split-GFP technology offers an affordable and technically simple manner for overcoming that constraints, making protein library screening more efficient through the normalization of the detected enzymatic activities in relation to the quantified protein contents responsible for them.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 41 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 41 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 14 34%
Student > Ph. D. Student 5 12%
Student > Bachelor 4 10%
Student > Master 3 7%
Other 2 5%
Other 4 10%
Unknown 9 22%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 17 41%
Agricultural and Biological Sciences 6 15%
Engineering 2 5%
Pharmacology, Toxicology and Pharmaceutical Science 1 2%
Unspecified 1 2%
Other 4 10%
Unknown 10 24%