| Autor: |
Pereira CF; Centre for Virology, Burnet Institute, Melbourne, Victoria, Australia., Ellenberg PC, Jones KL, Fernandez TL, Smyth RP, Hawkes DJ, Hijnen M, Vivet-Boudou V, Marquet R, Johnson I, Mak J |
| Jazyk: |
angličtina |
| Zdroj: |
PloS one [PLoS One] 2011 Feb 11; Vol. 6 (2), pp. e17016. Date of Electronic Publication: 2011 Feb 11. |
| DOI: |
10.1371/journal.pone.0017016 |
| Abstrakt: |
Due to its small size and versatility, the biarsenical-tetracysteine system is an attractive way to label viral proteins for live cell imaging. This study describes the genetic labeling of the human immunodeficiency virus type 1 (HIV-1) structural proteins (matrix, capsid and nucleocapsid), enzymes (protease, reverse transcriptase, RNAse H and integrase) and envelope glycoprotein 120 with a tetracysteine tag in the context of a full-length virus. We measure the impact of these modifications on the natural virus infection and, most importantly, present the first infectious HIV-1 construct containing a fluorescently-labeled nucleocapsid protein. Furthermore, due to the high background levels normally associated with the labeling of tetracysteine-tagged proteins we have also optimized a metabolic labeling system that produces infectious virus containing the natural envelope glycoproteins and specifically labeled tetracysteine-tagged proteins that can easily be detected after virus infection of T-lymphocytes. This approach can be adapted to other viral systems for the visualization of the interplay between virus and host cell during infection. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
|
