Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

Vol 19, Issue 4, 2017; Page No.(1034-1040)

INSERTION OF 2A ‘SELF-CLEAVING’ PEPTIDES BETWEEN ORF4 AND ORF5 OF GRAPEVINE VIRUS A GENOME FOR VECTOR DEVELOPMENT

DILYARA A. GRITSENKO, NURBOL N. GALIAKPAROV

Abstract

The development of new vectors based on genomes of plant viruses is actual goal for highefficiency protein expression in plants. In this work, grapevine virus A (GVA) has been utilized for vector creation. We have modified genome of GVA by insertion of 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 been subcloned 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 coagroinfiltration. This developed vectorcan be used for heterologous protein expression in plants by using magnifection method or by co-agroinfiltration with native virus.

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