Chapter title |
Bacterial Genome Editing Strategy for Control of Transcription and Protein Stability
|
---|---|
Chapter number | 3 |
Book title |
Synthetic Metabolic Pathways
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7295-1_3 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7294-4, 978-1-4939-7295-1
|
Authors |
Ida Lauritsen, Virginia Martínez, Carlotta Ronda, Alex Toftgaard Nielsen, Morten H. H. Nørholm |
Abstract |
In molecular biology and cell factory engineering, tools that enable control of protein production and stability are highly important. Here, we describe protocols for tagging genes in Escherichia coli allowing for inducible degradation and transcriptional control of any soluble protein of interest. The underlying molecular biology is based on the two cross-kingdom tools CRISPRi and the N-end rule for protein degradation. Genome editing is performed with the CRMAGE technology and randomization of the translational initiation region minimizes the polar effects of tag insertion. The approach has previously been applied for targeting proteins originating from essential operon-located genes and has potential to serve as a universal synthetic biology tool. |
X Demographics
Geographical breakdown
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Unknown | 4 | 100% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 4 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 12 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 4 | 33% |
Student > Master | 2 | 17% |
Researcher | 2 | 17% |
Student > Doctoral Student | 1 | 8% |
Unspecified | 1 | 8% |
Other | 1 | 8% |
Unknown | 1 | 8% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 5 | 42% |
Agricultural and Biological Sciences | 3 | 25% |
Unspecified | 1 | 8% |
Chemical Engineering | 1 | 8% |
Immunology and Microbiology | 1 | 8% |
Other | 0 | 0% |
Unknown | 1 | 8% |