Immunomodulation der Kartoffel-Blattrollvirus (PLRV) Multiplikation via Expression virusspezifischer rekombinanter Antikörper in planta

The proteolytical activity of the non-structural protein P1 of Potato leafroll virus (PLRV) leads to the formation of a subset of P1 derivates including P1-C25 and the VPg. In order to study the relevance of P1 and/or its derivate P1-C25 to the viral life-cycle the VH- and VL-domains of a monoclonal antibody mAbP1-C25 raised against the carboxy terminus of P1 was cloned in order to obtain a single chain variable fragment scFvP1-C25 that was subsequently analysed. The resulting antibody was bacterially expressed in the single chain format and designated as scFvP1-C25. The engineered scFvP1-C25 displayed identical binding specificity of the parental monoclonal antibody mAbP1-C25 and could be used for immunodetection of recombinant and native P1 and P1-C25. In a transient expression approach co-transformation of an infectious PLRV cDNA clone and the antibody scFvP1-C25 in Nicotiana tabacum strongly reduces virus accumulation. Transgenic potato plants expressing scFvP1-C25 showed high levels of resistance following PLRV inoculation by viruliferous aphids. In conclusion, transient or stable expression of a single chain fragment variable engineered against the carboxy terminus of P1 greatly reduces PLRV accumulation indicating the importance of P1 and/or P1-C25 for the life cycle of PLRV. This is the first report studying PLRV gene function in vivo without genetic alteration of the viral genome and of plantibody-mediated reduction in virus accumulation to a luteovirus.

Grantor dc:publisher

Publikationsserver der RWTH Aachen University

Year dc:date

2007

Author dc:creator

• Nickel, Holger

Contributors dc:contributor

• Fischer, Rainer

• info:eu-repo/classification/ddc/570
• Biotechnologie
• Pflanzenkrankheit
• Biowissenschaften, Biologie
• molekulare Pflanzenbiotechnologie
• biotechnology
• molecular plant biotechnology

Statement dc:rights

• info:eu-repo/semantics/openAccess

Language dc:language

ger

Identifier

https://publications.rwth-aachen.de/search?p=id:%22RWTH-CONV-112791%22

OAI record oai:identifier

https://publications.rwth-aachen.de/oai2d?verb=GetRecord&metadataPrefix=oai_dc&identifier=oai%3Apublications.rwth-aachen.de%3A50238