Small Functional Peptides and Their Application in Superfunctionalizing Proteins

Billerbeck, Sonja

doi:10.1002/9783527688104.ch11

Cited by 2 publications

(1 citation statement)

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“…Besides changing single amino acids, several protein engineering approaches require the insertion of short stretches of DNA that either encode for functional peptide tags [55] – such as affinity tags for purification or detection, [56] protease cleavage sites for on‐demand inactivation,[ 57 , 58 ] tags for post‐translational modification via click chemistry, [59] or labeling sites for protein visualization [60] – or encode for molecular recognition sites required for further library processing; for example, restriction enzymes sites that can subsequently be used for creating systematic insertion or deletion libraries.…”

Section: Encoding Short Dna Stretches: Oligo Pool‐based Insertion And...mentioning

confidence: 99%

Oligo Pools as an Affordable Source of Synthetic DNA for Cost‐Effective Library Construction in Protein‐ and Metabolic Pathway Engineering

2021

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The construction of custom libraries is critical for rational protein engineering and directed evolution. Array-synthesized oligo pools of thousands of user-defined sequences (up to ~350 bases in length) have emerged as a low-cost commercially available source of DNA. These pools cost � 10 % (depending on error rate and length) of other commercial sources of custom DNA, and this significant cost difference can determine whether an enzyme engineering project can be realized on a given research budget. However, while being cheap, oligo pools do suffer from a low concentration of individual oligos and relatively high error rates. Several powerful techniques that specifically make use of oligo pools have been developed and proven valuable or even essential for next-generation protein and pathway engineering strategies, such as sequence-function mapping, enzyme minimization, or de-novo design. Here we consolidate the knowledge on these techniques and their applications to facilitate the use of oligo pools within the protein engineering community.

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Section: Encoding Short Dna Stretches: Oligo Pool‐based Insertion And...mentioning

confidence: 99%

Oligo Pools as an Affordable Source of Synthetic DNA for Cost‐Effective Library Construction in Protein‐ and Metabolic Pathway Engineering

2021

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Combining Oligo Pools and Golden Gate Cloning to Create Protein Variant Libraries or Guide RNA Libraries for CRISPR Applications

Valero,

Prins,

de Vroet

et al. 2024

Methods in Molecular Biology

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Small Functional Peptides and Their Application in Superfunctionalizing Proteins

Cited by 2 publications

References 123 publications

Oligo Pools as an Affordable Source of Synthetic DNA for Cost‐Effective Library Construction in Protein‐ and Metabolic Pathway Engineering

Oligo Pools as an Affordable Source of Synthetic DNA for Cost‐Effective Library Construction in Protein‐ and Metabolic Pathway Engineering

Combining Oligo Pools and Golden Gate Cloning to Create Protein Variant Libraries or Guide RNA Libraries for CRISPR Applications

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