M.Sc. Tezi Görüntüleme | |||||||||||||||||||||
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| Summary: SUMMARY Study towards knockdown of Spodoptera exigua pkg gene by antisense RNA
As a result of infection, some behavioural changes occur in insects infected with baculoviruses. These behavioural changes include hypermobility and climbing the top of the plant. The mechanisms, by which the virus infection induces this behavioural change, are largely unknown. In this study, the role of cGMP protein kinase (pkg) gene thought to be responsible from behavioural changes occurred in host induced by the virus have been examined. It is shown that for gene provided food searching context and mobility in different insects, such as Drosophila melanogaster and Apis mellifera. The aim of the study is to silence Spodoptera exigua pkg gene and determine the role of pkg gene in the behavioural response of the host. In the study initially, it is detected that pkg and gfp genes were down regulated with AcMNPV bacmid producing dsRNA against these genes. For this purpose, AcMNPV bacmids expressing dsRNAs against target genes (pkg and gfp) separately co-infected with recombinant AcMNPV expressing gfp to Sf-9 cells. As a result of the infection, it was determined that pkg and gfp genes were successfully down regulated in the 3 d.p.i and the 4 d.p.i, respectively. Following these experiments, in order to test different systems for down regulating pkg gene, the plasmid vectors necessary for constructing bacmid producing dsRNA at hairpin structure against pkg gene.In this study it is aimed to design the technology that will be used for the production of viral bacmid system potentially useful for understanding the mechanism involving hypermobility behaviour in host insects larvae’s upon viral infection Key Words: Baculoviruses, Spodoptera exigua, cGMP protein kinase (pkg) gene, RNA interference
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