| |  Online-Ressource |
|---|
| Verfasst von: | Füllgrabe, Marc W. [VerfasserIn]  |
|---|
| | Boonrod, Kajohn [VerfasserIn] |
|---|
| | Jamous, Rana [VerfasserIn] |
|---|
| | Moser, Mirko [VerfasserIn] |
|---|
| | Shiboleth, Yoel [VerfasserIn] |
|---|
| | Krczal, Gabi [VerfasserIn] |
|---|
| | Wassenegger, Michael [VerfasserIn] |
|---|
| Titel: | Expression, purification and functional characterization of recombinant Zucchini yellow mosaic virus HC-Pro |
|---|
| Verf.angabe: | Marc W. Fuellgrabe, Kajohn Boonrod, Rana Jamous, Mirko Moser, Yoel Shiboleth, Gabi Krczal, Michael Wassenegger |
|---|
| Jahr: | 2011 |
|---|
| Umfang: | 6 S. |
|---|
| Fussnoten: | Gesehen am 31.05.2023 ; Online verfügbar 30 Juli 2010 |
|---|
| Titel Quelle: | Enthalten in: Protein expression and purification |
|---|
| Ort Quelle: | Amsterdam [u.a.] : Elsevier, 1990 |
|---|
| Jahr Quelle: | 2011 |
|---|
| Band/Heft Quelle: | 75(2011), 1 vom: Jan., Seite 40-45 |
|---|
| ISSN Quelle: | 1096-0279 |
|---|
| Abstract: | HC-Pro is a helper component-proteinase which acts as a multifunctional protein in the potyviral life cycle. Apart from its proteolytic activity, HC-Pro has the capacity to bind duplex small RNAs (sRNAs). To investigate HC-Pro-mediated sRNA binding in vitro, high amounts of purified protein are required. For this purpose, the Zucchini yellow mosaic virus (ZYMV) HC-Pro was expressed as a fusion with hexa-histidine (6xHis) or maltose-binding protein (MBP) in Escherichia coli. The expressed fusion proteins were purified by affinity chromatography. 6xHis:HC-Pro and MBP:HC-Pro were partially soluble. Electrophoretic mobility-shift assays demonstrated that only MBP:HC-Pro exhibits the sRNA binding activity. The recombinant HC-Pro bound 21bp siRNAs as well as 19bp and 24bp siRNAs. A point mutation in the highly conserved FRNK box produced the HC-ProFINK protein, previously shown to be associated with reduced viral symptoms and weak sRNA binding. In this study, sRNA binding of the MBP:HA-HC-ProFINK was not detectable. The high yield of purified HC-Pro offers the possibility to study the biochemistry of the protein in detail. |
|---|
| DOI: | doi:10.1016/j.pep.2010.07.008 |
|---|
| URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
Volltext: https://doi.org/10.1016/j.pep.2010.07.008 |
|---|
| | Volltext: https://www.sciencedirect.com/science/article/pii/S1046592810002135 |
|---|
| | DOI: https://doi.org/10.1016/j.pep.2010.07.008 |
|---|
| Datenträger: | Online-Ressource |
|---|
| Sprache: | eng |
|---|
| Sach-SW: | HC-Pro |
|---|
| | Potyviruses |
|---|
| | sRNA binding |
|---|
| | ZYMV |
|---|
| K10plus-PPN: | 1847023932 |
|---|
| Verknüpfungen: | → Zeitschrift |
|---|
Expression, purification and functional characterization of recombinant Zucchini yellow mosaic virus HC-Pro / Füllgrabe, Marc W. [VerfasserIn]; 2011 (Online-Ressource)
