Chapter title |
Selection of Highly Expressed Gene Variants in Escherichia coli Using Translationally Coupled Antibiotic Selection Markers
|
---|---|
Chapter number | 16 |
Book title |
Synthetic Metabolic Pathways
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7295-1_16 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7294-4, 978-1-4939-7295-1
|
Authors |
Maja Rennig, Daniel O. Daley, Morten H. H. Nørholm |
Abstract |
Strategies to select highly expressed variants of a protein coding sequence are usually based on trial-and-error approaches, which are time-consuming and expensive. We address this problem using translationally coupled antibiotic resistance markers. The system requires that the target gene can be fused at the 3'-end with a translational coupling element and an antibiotic resistance gene. Highly expressed target genes can then be selected using a fast and simple whole cell survival assay in the presence of high antibiotic concentrations. Herein we show that the system can be used to select highly expressing clones from libraries sampling translation initiation sites. |
X Demographics
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United Kingdom | 1 | 100% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 16 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 3 | 19% |
Student > Master | 3 | 19% |
Student > Bachelor | 2 | 13% |
Researcher | 1 | 6% |
Unknown | 7 | 44% |
Readers by discipline | Count | As % |
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Agricultural and Biological Sciences | 4 | 25% |
Biochemistry, Genetics and Molecular Biology | 3 | 19% |
Immunology and Microbiology | 1 | 6% |
Unknown | 8 | 50% |