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kwedul

A Multicomponent Animal Virus Isolated from Mosquitoes: Cell Host & Microbe - 68 views

www.cell.com/...S1931-3128(16)30310-9

shared by kwedul on 02 Sep 16 - Cached
  • reverse genetics system
    • Casey Finnerty
      Casey Finnerty on 02 Sep 16
       
      Please be ready to explain why reverse genetics was a necessary approach for studying this virus.
  • GCXV
    • Casey Finnerty
      Casey Finnerty on 14 Sep 16
       
      Here is an example of a stickie note.
  • Viral replication was detected in three mosquito cell lines and in intrathoracically inoculated adult female mosquitoes
  • ...14 more annotations...
  • Selective degradation with RNase I demonstrates that GCXV has a single-stranded RNA genome.
    • Casey Finnerty
      Casey Finnerty on 19 Sep 16
       
      RNaseI is specific for ssRNA.
  • reverse genetics system
  • Multicomponent plant viruses were recognized on the basis of deviations from the expected relationship between infectious dose and the number of lesions on infected leaves (i.e., exhibiting multi-hit rather than single-hit kinetics; Flint et al., 2009xPrinciples of Virology Volume I: Molecular Biology. Flint, S.J., Enquist, L.W., Racaniello, V.R., Skalka, A.M., Barnum, D.R., and de Evaluación, E. See all References, Sánchez-Navarro et al., 2013xEffects of the number of genome segments on primary and systemic infections with a multipartite plant RNA virus. Sánchez-Navarro, J.A., Zwart, M.P., and Elena, S.F. J. Virol. 2013; 87: 10805–10815Crossref | PubMedSee all References).
    • jsackett13
      jsackett13 on 19 Sep 16
       
      This was explained in the blog article.
  • of any other group of organisms (
    • aceastep
      aceastep on 19 Sep 16
       
      wow
  • no Illumina reads aligned to the segment 5 sequences from the other isolates, and no contigs from ACH27 exhibited significant similarity to the segment 5 sequences from the other isolates. Therefore, segment 5 appears to be absent from ACH27 and TR7094.
  • However, only four segments were assembled for ACH27 and TR7094 (genome size ∼10.6 kb). For these two isolates, the four assembled segments corresponded to the four largest segments assembled in the other isolates (Figure 1Figure 1, segments 1–4).
  • The phylogeny inferred from segment 5 is also inconsistent with those of the other segments. In addition to the absence of this segment in TR7094 and ACH27, this segment exhibited very low levels of nt divergence (0.4%–2.3%), with most sequence variations only present in a single isolate.
  • with each genome segment separately packaged into virions. Although multicomponent genomes are relatively common among RNA viruses that infect plants and fungi, this method of genome organization has not previously been seen in animal viruses (
    • kwedul
      kwedul on 21 Sep 16
       
      Is this the only thing required for a genome to be considered multicomponent?
  • RNA extracted from purified GCXV particles confirmed the presence of a segmented genome (
    • kwedul
      kwedul on 21 Sep 16
       
      So what exactly is it about this purified GCXV that shows it is segmented?
  • The mosquito pools for both ACH27 and TR7094 contained multiple viruses capable of replicating in mosquito cells, so we were unable to obtain pure cultures for these isolates (
    • kwedul
      kwedul on 21 Sep 16
       
      What is it about there being multiple viruses capable of replicating that made them unable to obtain pure cultures for the isolates?
  • Nuclease digestion assays (Figure 1Figure 1B), along with 5′ and 3′ rapid amplification of cDNA ends (RACE), confirmed that all five genome segments were single-stranded, positive-sense RNA (ssRNA+).
    • kwedul
      kwedul on 21 Sep 16
       
      What about these tests confirms all five genome segments were single-stranded positive-sense RNA?
  • all five segments, (2) segments 1–4, and (3) segments 2–5
    • kwedul
      kwedul on 21 Sep 16
       
      Why did they use these three combinations?
  • the number of plaques decreased more quickly than expected with dilution of the inoculant) (
    • kwedul
      kwedul on 21 Sep 16
       
      Why? What is the significance of this?
  • segments 1–5 (passage 1) and segments 1–4 (passage 2)
    • kwedul
      kwedul on 21 Sep 16
       
      What about a second passage, including 1-4, makes it brighter?
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