ИСТИНА |
Войти в систему Регистрация |
|
ИСТИНА ИНХС РАН |
||
Potato virus X (PVX) (helical filamentous plant virus, genus Potexvirus, family Alphaflexiviridae) has the possibility of reversible dissociation of virions into coat proteins (CP) and RNA followed by the in vitro self-assembly of viral particles which save the structure and biological activity of the virus. Previously we showed that in vitro incubation of PVX CP with the RNAs of various plant and animal viruses results in the viral ribonucleoproteins (vRNP) formation (Arkhipenko et al., 2011). The helical structure, morphology and translational properties of these artificial vRNP were identical to cognate vRNP assembled from PVX RNA and PVX CP and to PVX virions. However, the widely used methods do not allow us to estimate the efficiency of assembly of artificial vRNPs formed by PVX CP and different plant and animal RNAs. To obtain data about the particles size (hydrodynamic diameter) and concentration the method of Nanoparticle Tracking Analysis (NTA) was used. PVX CP was incubated with PVX RNA and different viral RNAs. Formed cognate and artificial vRNP were analyzed by NTA. This method based on laser-illuminated optical microscopy enables to detect the RNPs assembly in real-time in liquid. The detected number of formed particles indicates the assembly efficiency. For the first time the information about efficiency of the potato virus X particles assembly was obtained by NTA. The assembly efficiency of RNPs formed by PVX CP with PVX RNA and heterologous viruses RNA was compared. The proposed method allows studying the mechanisms of initiation and elongation of the viral ribonucleoproteins. Reference: 1. Arkhipenko et al., Acta naturae 2011, 3, № 3(10), 40-46.