| Chapter title |
Reverse genetic engineering of the human rhinovirus serotype 16 genome to introduce an antibody-detectable tag.
|
|---|---|
| Chapter number | 13 |
| Book title |
Rhinoviruses
|
| Published in |
Methods in molecular biology, January 2015
|
| DOI | 10.1007/978-1-4939-1571-2_13 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-1570-5, 978-1-4939-1571-2
|
| Authors |
Erin J Walker, Lora M Jensen, Reena Ghildyal, Erin J. Walker, Lora M. Jensen |
| Abstract |
The ability to accurately detect viral proteins during infection is essential for virology research, and the lack of specific antibodies can make this detection difficult. Reverse genetic engineering of virus genomes to alter the wild-type genome is a powerful technique to introduce a detectable tag onto a viral protein. Here we outline a method to incorporate an influenza hemagglutinin epitope tag onto the 2A protease of HRV16. The method uses site-directed mutagenesis PCR to introduce the sequence for the HA antigen onto either the C or N termini of 2A protease while keeping the relevant internal cleavage sites intact. The new viral product is then cloned into a wild-type HRV16 plasmid and transfected into Ohio Hela cells to produce recombinant virus. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Australia | 1 | 50% |
| Unknown | 1 | 50% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 1 | 50% |
| Members of the public | 1 | 50% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 9 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 2 | 22% |
| Student > Master | 2 | 22% |
| Other | 1 | 11% |
| Student > Bachelor | 1 | 11% |
| Professor > Associate Professor | 1 | 11% |
| Other | 1 | 11% |
| Unknown | 1 | 11% |
| Readers by discipline | Count | As % |
|---|---|---|
| Immunology and Microbiology | 2 | 22% |
| Agricultural and Biological Sciences | 2 | 22% |
| Biochemistry, Genetics and Molecular Biology | 1 | 11% |
| Business, Management and Accounting | 1 | 11% |
| Computer Science | 1 | 11% |
| Other | 1 | 11% |
| Unknown | 1 | 11% |