| Chapter title |
Preparation of Recombinant Alphaviruses for Functional Studies of ADP-Ribosylation
|
|---|---|
| Chapter number | 21 |
| Book title |
ADP-ribosylation and NAD+ Utilizing Enzymes
|
| Published in |
Methods in molecular biology, August 2018
|
| DOI | 10.1007/978-1-4939-8588-3_21 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-8587-6, 978-1-4939-8588-3
|
| Authors |
Rachy Abraham, Robert Lyle McPherson, Easwaran Sreekumar, Anthony K. L. Leung, Diane E. Griffin, Abraham, Rachy, McPherson, Robert Lyle, Sreekumar, Easwaran, Leung, Anthony K. L., Griffin, Diane E. |
| Abstract |
Recently we characterized the mono(ADP-ribosyl) hydrolase (MAR hydrolase) activity of the macrodomain of nonstructural protein 3 (nsP3MD) of chikungunya virus. Using recombinant viruses with targeted mutations in the macrodomain, we demonstrated that hydrolase function is important for viral replication in cultured neuronal cells and for neurovirulence in mice. Here, we describe the general cell culture and animal model infection protocols for alphaviruses and the technical details for biochemical characterization of the MAR hydrolase activity of nsP3MD mutants and the preparation of recombinant viruses incorporating those mutations through site-directed mutagenesis of an infectious cDNA virus clone. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 15 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 3 | 20% |
| Student > Doctoral Student | 2 | 13% |
| Student > Postgraduate | 2 | 13% |
| Researcher | 2 | 13% |
| Student > Ph. D. Student | 1 | 7% |
| Other | 2 | 13% |
| Unknown | 3 | 20% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 5 | 33% |
| Biochemistry, Genetics and Molecular Biology | 2 | 13% |
| Arts and Humanities | 1 | 7% |
| Immunology and Microbiology | 1 | 7% |
| Social Sciences | 1 | 7% |
| Other | 1 | 7% |
| Unknown | 4 | 27% |