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Virol Res J 2017 Volume 1 Issue 3
International Virology Conference
October 30-31, 2017 | Toronto, Canada
Modification of grapevine virus A genome for vector development
Gritsenko D A
1, 2
, Deryabina N D
1
and
Galiakparov N N
1
1
Institute of Plant Biology and Biotechnology, Kazakhstan
2
Al-Farabi Kazakh National University, Kazakhstan
T
he development of new vectors based on plant viruses
does not discontinue for the goal of creation a more
efficient vector with high expression of heterologous proteins.
We attempted to develop such vector based on genome of
grapevine virus A (GVA). For creation of this viral vector we
inserted gene encoding coat protein of Apple chlorotic leafspot
virus (ACLSV) before ORF4 of GVA. PCASS vector carrying the
complete genome of the grapevine virus A (pCASSgva) was
used for creation a viral vector based on GVA. The viral genome
was modified by introducing a CP gene of ACLSV before ORF 4
within restriction sites XmaI and XbaI. The overlapping region
of 3’- terminus ORF3 and 5’- terminus of ORF4 was intact. The
CP gene of ACLSV was placed under control of CP subgenomic
promoter of GVA. The modified viral genome was subcloned
into a pCambia 2300 binary vector. The expression of the CP
of ACLSV in agroinfiltrated
N.benthamiana
leaves after 3-4
days of infection was confirmed by using western blotting.
Agroinfiltration of transgenic plants carrying CP of GVA was not
successful, we assume due RNA-silencing. It will be investigated
the expression level of the viral vector and its usefulness as a
vector for the expression of avian influenza hemagglutinin.
Speaker Biography
Gritsenko D A is a PhD- student at Kazakh National University named after al- Farabi.
She performs her diploma work at Institute of Plant Biology and Biotechnology. The
title of diploma is “Development of Viral Vector for Heterologouos Protein Expression
in Plants”. She developed 2 vectors based on genome of Grapevine virus A by using
main strategies for vector engineering such as “deconstructed virus” and “full virus”.
Currently, these vectors were investigated for successful expression of eGFP and coat
protein of Apple chlorotic leafspot virus. Moreover, she developed transgenic plants
carrying coat protein of GVA for increasing of target protein yield since GVA cannot
move between cells in non-encapsidated form.
e:
d.kopytina@gmail.com