| Chapter title |
A Rapid Method for Engineering Recombinant Polioviruses or Other Enteroviruses
|
|---|---|
| Chapter number | 13 |
| Book title |
Poliovirus
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3292-4_13 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3291-7, 978-1-4939-3292-4
|
| Authors |
Maël Bessaud, Isabelle Pelletier, Bruno Blondel, Francis Delpeyroux, Bessaud, Maël, Pelletier, Isabelle, Blondel, Bruno, Delpeyroux, Francis |
| Abstract |
The cloning of large enterovirus RNA sequences is labor-intensive because of the frequent instability in bacteria of plasmidic vectors containing the corresponding cDNAs. In order to circumvent this issue we have developed a PCR-based method that allows the generation of highly modified or chimeric full-length enterovirus genomes. This method relies on fusion PCR which enables the concatenation of several overlapping cDNA amplicons produced separately. A T7 promoter sequence added upstream the fusion PCR products allows its transcription into infectious genomic RNAs directly in transfected cells constitutively expressing the phage T7 RNA polymerase. This method permits the rapid recovery of modified viruses that can be subsequently amplified on adequate cell-lines. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 9 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 2 | 22% |
| Student > Bachelor | 2 | 22% |
| Student > Doctoral Student | 1 | 11% |
| Researcher | 1 | 11% |
| Professor > Associate Professor | 1 | 11% |
| Other | 0 | 0% |
| Unknown | 2 | 22% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 2 | 22% |
| Medicine and Dentistry | 2 | 22% |
| Biochemistry, Genetics and Molecular Biology | 1 | 11% |
| Immunology and Microbiology | 1 | 11% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 11% |
| Other | 0 | 0% |
| Unknown | 2 | 22% |