TESIS NOEMI HERRERO.pdf - Consejo Superior de Investigaciones ...

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Capítulo III
Fig. 4. Alignment of the 3’ UTR sequences of eleven members of the Victorivirus genus and TcV1. A
conserved sequence (blue letters) among members of this genus is indicated.
Analysis of virus particles and structural proteins
Partially purified virions from strains 11 and 3398M were disrupted by treatment
with SDS and phenol in order to determine if all dsRNAs infecting these strains were
encapsidated. All dsRNAs infecting each strain were recovered from the aqueous phase
obtained after the disruption treatment. This result suggests that all six dsRNA elements
are encapsidated in protein particles.
When a sucrose gradient purified virus preparation of strain 11-1L, harbouring only
dsRNA1, was examined by transmission electron microscopy (TEM), isometric virus like
particles of approximately 50 nm of diameter were observed (Fig. 5).
Fig. 5. A. SDS-PAGE analysis of purified TcV1 particles. Lane M, molecular weight marker (kDa); lane 1,
SDS-treated TcV1 virions. The structural proteins were visualized by coomassie blue staining. The 79.7 kDa
protein purified by electrophoresis (CP) has a molecular weight similar to the one estimated for the
hypothetical protein coded by ORF1 (79.87 kDa). B. Isometric virus-like particles observed by TEM from a
purified virus preparation from T. cylindrosporum 11-1L isolate harbouring only dsRNA1. Bar, 25 nm.
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