INSERTION OF 2A 'SELF-CLEAVING' PEPTIDES BETWEEN ORF4 AND ORF5 OF GRAPEVINE VIRUS A GENOME FOR VECTORDEVELOPMENT

摘要

The development of new vectors based on genomes of plant viruses is actual goal for high-efficiency protein expression in plants. In this work, grapevine virus A (GVA) has been utilized for vector creation. We have modified genome of GVA by insertionof 2A peptides between ORF 4 and ORF5. Moreover, coat protein gene of ACLSV (apple chlorotic leaf spot virus) has been introduced between 2A peptides for investigation of heterologous gene expression efficiency in plants. Modified genome of GVA has beensubcloned into binary vector and has been delivered to plants via agroinfiltration. Expression of ACLSV CP was confirmed in infiltrated leaves on 3 day and in upper leaves on 10 day after co-agroinfiltration. This developed vectorcan be used for heterologous protein expression in plants by using magnifection method or by co-agroinfiltration with native virus.