Gene expression profile of iridovirus-infected orange-spotted grouper Epinephelus coioides kidney
Date Issued
2008
Date
2008
Author(s)
Huang, Hseng-Hsiang
Abstract
Grouper iridovirus (GIV), a double strand DNA virus, encoded 139 open reading framesORFs) in its 140 kb genome size. Iridovirus mainly attack immune system such as spleen andidney of host. The research of virus gene expression profile upon infection had been revealed.owever, investigation of host gene expression during virus infection is still unknown. In order tonderstand virus infection mechanism and gene expressing profile of host, we have constructedIV-infected grouper kidney λcDNA library. After transfering the vector from phage to plasmid,otal 6000 candidate plasmids were purified and spotted on to cDNA microarray chip.esulting chip was hybridized with different time-coursed fluorsence-labeled cDNA probesade from GIV, polyriboinosinic polyribocytidylic (Poly I:C) or lipopolysaccharide (LPS)-injectedrouper larvae kidney total RNA. Images obtained from dual-laser scanner were analyzed andenes were selected by the criteria of at least 2.5-fold up or down-regulation of expression level inny of the experiment days. Candidate genes were sequenced, translated into protein sequence andlasted in NCBI protein database. Total 53 non-redundant genes were obtained, in which 22 ofhem were up-regulated and the rest 31 genes were down-regulated. Functional categories referredhese genes into 11 groups, which were apoptosis, ubiquitin/protein degradation, respiratory chain,mmune, transcription, translation, structure protein, erythropoiesis, proton pump, metabolism andenes with unknown function.elected genes from this experiment help us elucidating the strategies of host genes inesponse to virus propagation and host defense during virus infection. Meanwhile, the plasmidNA chip is proof of use in searching host differential expressed genes.
Subjects
iridovirus
microarray
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