plant virology literature review
shiva charan
ms_thesis.pptxStudy on the role of GTP Binding Protein of the Infected Plants in PVA Infection by Virus Induced Gene Silencing Ras-related protein Rab1a
OBJECTIVES
To examine the role of GTP binding protein which was only found in PVA infected nuclei, in PVA infection by virus induced gene silencing by comparing infected and un-infected plants in Nicotiana benthamiana
To check the RNA levels in the inoculated leaves using quantitative real time PCR.
No silencing found in the controlled and infected plants.
Gene silencing (Rab1a) doesnt or have negligible effect on the poty viral infection.
GTP binding ras protein may help in the viral infection which will be proved by the gene silencing and qPCR.
HYPOTHESIS
GTP binding proteins are a set of family proteins which are very important in various metabolic processes.
They are also associated with nuclear-cytoplasmic trafficking
A recent study showed that a small specific GTP binding protein is found in the nucleus of PVA infected potato plant, whereas there is no proteins in the control plant.
When this protein sequence is searched for the similarity, it is found that there is a similar protein in Nicotiana benthamiana.
In this study, we examine the role of this small protein in PVA infection.
METHODS
RNA EXTRACTION & cDNAsynthesis
PCR AMPLIFICATION
RESTRICTION DIGESTION , LIGATION AND TRANSFORMATION
COLONY PCR
PLASMID ISOLATION AND DNA SEQUENCING
AGRO TRANSFORMATION (GV3101 + psoup)
PRESENT WORK
Growing plants
Constructs preparation:
i) GV3101+PDS
ii) GV3101+empty
iii) GV3101+ insert
iv) GV3101+PVA-gfp
Infiltration
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