Molecular characterisation of the sgRNAs of GLRaV-3


Project Number
GENUS 07-03

Project title
Molecular characterisation of the sgRNAs of GLRaV-3

Project leader
Burger, J T

Institution
University of Stellenbosch. Department of Genetics

Team members
Burger, J T
Freeborough, M J
Maree, H J

Project description
The main aim of this project is to address the lack of knowledge that exists on the replication strategy of grapevine leafroll associated virus 3 (GLRaV-3). Very little is known about the role of GLRaV-3 sgRNAs other than that it forms part of virus replication and viral gene regulation. This project will investigate the production and composition of the sgRNAs of GLRaV-3 with particular emphasis on the sg-promoters.

They are of particular interest as they are very strong viral promoters that could possibly be utilized to combat leafroll disease. The sg-promoter requires the replication proteins of GLRaV-3 (ORF1a and b) to be functional and can thus be used for virus-induced gene expression. GM plants containing this construct will only express an RNA molecule of the target gene in the negative orientation. Upon virus infection the negative sense will be transcribed with the RdRp into a positive sense mRNA of the target gene that can be translated into the required gene product. This approach limits the metabolic load and associated problems found with constitutive expression. Developing new promoters (synthetic or native) derived from the sg-promoters of GLRaV-3 minimise the amount of licensing required when implementing the technology for commercial applications.

Poster(s)
Maree, H J, Freeborough, M J and Burger, J T. 2008. Characterisation of the grapevine leafroll-associated virus 3 replication mechanism. Poster presented at the Agricultural Biotechnology International Conference. 24-27 August, Cork, Ireland.

Coetzee, B, Maree, H J, Jooste, A E C, Freeborough, M J and Burger, J T. 2008. Comparison of the genetic variation of three regions in the GLRaV-3 genome. Poster presented at the 31st National Conference of the South African Society for Enology and Viticulture. 11-14 November, Somerset West, South Africa

Presentation(s)
Maree, H J, Freeborough, M-J and Burger, J T. 2008. Characterisation of the grapevine leafroll-associated virus 3 sgRNAs. Paper presented at the at the Cape Biotechnology Forum. 30 November – 2 December, Somerset West, South Africa.

Maree, H J, Jooste, A E C, Stephan, D, Freeborough, M J and Burger, J T. 2009. Characterisation of the genomic and subgenomic RNA of Grapevine leafroll associated virus-3 (GLRaV-3). Paper presented at the 16th Meeting of the International Council for the Study of Virus and Virus-like Diseases of the Grapevine. 31 August – 4 September, Dijon, France.

Maree, H J, Freeborough, M J and Burger, J T. 2009. Characterisation of the Grapevine leafroll-associated virus sgRNAs. Paper presented at the 46th Congress of the South African Society for Plant Pathology and the 6th Congress of the African Mycological Association. 25-28 January, Gordon’s Bay, South Africa.

Article
Maree, H J, Freeborough, M J, Burger, J T. 2008. Complete nucleotide sequence of a South African isolate of grapevine leafroll-associated virus 3 reveals a 5’UTR of 737 nucleotides, Archives of Virology, Mnth Apr v. 153 (4) (p. 755-757)

Maree, H J, Gardner, H F J, Freeborough, M J, Burger, J T. 2010. Mapping of the 5′ terminal nucleotides of grapevine leafroll-associated virus 3 sgRNAs, Virus Research, v. 151 (p. 252-255)

FinalReport.pdf

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