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TitleAffimer reagents as tools in diagnosing plant virus diseases.
Journal, issue, pagesSci Rep, Vol. 9, Issue 1, Page 7524, Year 2019
Publish dateMay 17, 2019
AuthorsEmma L Hesketh / Christian Tiede / Hope Adamson / Thomas L Adams / Matthew J Byrne / Yulia Meshcheriakova / Inga Kruse / Michael J McPherson / George P Lomonossoff / Darren C Tomlinson / Neil A Ranson /
PubMed AbstractPlant viruses can cause devastating losses to agriculture and are therefore a major threat to food security. The rapid identification of virally-infected crops allowing containment is essential to ...Plant viruses can cause devastating losses to agriculture and are therefore a major threat to food security. The rapid identification of virally-infected crops allowing containment is essential to limit such threats, but plant viral diseases can be extremely challenging to diagnose. An ideal method for plant virus diagnosis would be a device which can be implemented easily in the field. Such devices require a binding reagent that is specific for the virus of interest. We chose to investigate the use of Affimer reagents, artificial binding proteins and a model plant virus Cowpea Mosaic virus (CPMV) empty virus like particles (eVLPs). CPMV-eVLP mimic the morphology of wild-type (WT) CPMV but lack any infectious genomic material and so do not have biocontainment issues. We have produced and purified an Affimer reagent selected for its ability to bind to CPMV-eVLP and have shown that the selected Affimer also specifically binds to WT CPMV. We have produced a 3.4 Å structure of WT CPMV bound to the Affimer using cryo-electron microscopy. Finally, we have shown that this Affimer is capable of reliably detecting the virus in crude extracts of CPMV-infected leaves and can therefore form the basis for the future development of diagnostic tests.
External linksSci Rep / PubMed:31101847 / PubMed Central
MethodsEM (single particle)
Resolution3.4 Å
Structure data

4610

6qoz

EMDB-4610: CryoEM reconstructuion of Cowpea Mosaic Virus (CPMV) bound to an Affimer reagent
PDB-6qoz: CryoEM reconstruction of Cowpea Mosaic Virus (CPMV) bound to an Affimer reagent
Method: EM (single particle) / Resolution: 3.4 Å

Source
  • synthetic construct (others)
  • cowpea mosaic virus
KeywordsVIRUS / CPMV / COMOVIRIDAE / PICORNAVIRALES / AFFIMEr REAGENT / antibody-like

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