| Chapter title |
Codon Optimizing for Increased Membrane Protein Production: A Minimalist Approach.
|
|---|---|
| Chapter number | 4 |
| Book title |
Heterologous Expression of Membrane Proteins
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3637-3_4 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3635-9, 978-1-4939-3637-3
|
| Authors |
Kiavash Mirzadeh, Stephen Toddo, Morten H. H. Nørholm, Daniel O. Daley |
| Editors |
Isabelle Mus-Veteau |
| Abstract |
Reengineering a gene with synonymous codons is a popular approach for increasing production levels of recombinant proteins. Here we present a minimalist alternative to this method, which samples synonymous codons only at the second and third positions rather than the entire coding sequence. As demonstrated with two membrane-embedded transporters in Escherichia coli, the method was more effective than optimizing the entire coding sequence. The method we present is PCR based and requires three simple steps: (1) the design of two PCR primers, one of which is degenerate; (2) the amplification of a mini-library by PCR; and (3) screening for high-expressing clones. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 12 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 4 | 33% |
| Student > Ph. D. Student | 3 | 25% |
| Student > Bachelor | 2 | 17% |
| Unknown | 3 | 25% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 4 | 33% |
| Agricultural and Biological Sciences | 4 | 33% |
| Neuroscience | 1 | 8% |
| Chemistry | 1 | 8% |
| Unknown | 2 | 17% |