Molecular characterization, expression and functional assay of grouper iridovirus 027L gene
Date Issued
2010
Date
2010
Author(s)
Shih, Chueh-Ju
Abstract
Groupers are vital fish in Taiwan, while grouper iridovirus (GIV) has caused mass mortality, which economically impacts on culture of marine fish. Iridovirus infections have led to serious economic loss in the aquaculture industry in Southern Taiwan. Our laboratory had determined the complete DNA sequence of GIV and identified 21 immediate-early genes from GIV by using the microarray method, which include GIV 027L open reading frame (ORF).
This GIV027L ORF is a caspase recruitment domain (CARD) domain containing gene which encodes a protein of 91 amino acid with predicted molecular mass of 10,505 Da and shows high similarity to human CARD-only protein, ICEBERG and pseudo-ICE.
RT-PCR result revealed that GIV027L transcript at an early stage (1 h) of GIV infection. Cycloheximide and aphidicolin further confirmed that GIV027L was an immediate-early gene. By using E. coli pET20b(+) expression system, the recombinant GIV027L protein can be used to immunize New Zealand rabbits to produce rabbit anti-GIV027L polyclonal antibody. The subcellular localization of pEGFP-N/GIV027L fusion protein was in the nucleus as well as cytoplasm. By immunocytochemical staining, it was proven that GIV027L-expressing cells can inhibit apoptosis induced by UV irridiation.
Subjects
grouper
iridovirus
Type
thesis
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