White Spot Syndrome Virus (WSSV) is the most devastating viral disease affecting cultivated shrimp around
the world. The CENIACUA breeding programme included host resistance as strategy to control WSSV. Since
1999, all families were subjected to WSSV challenge tests and up to 5% of the animals survived. We suspect
that a unique genetic profile could be responsible for survival. By Subtractive Suppression Hybridization
improved with Mirror Orientation Selection we compared gene expression in haemocytes of WSSV-infected
shrimp; seven were previously challenged (pre-challenged) and 10 naïve. Out of 1167 clones, 993 (85%) were
obtained from the WSSV-infected pre-challenged library, while 174 (15%) were from the WSSV-infected naïve
library. Differential expression was confirmed by hybridization using PCR DIG-labelled probes from
unsubtracted cDNA. The 216 (18.5%) clones differentially expressed in the WSSV-infected pre-challenged
library and 23 (2%) in the WSSV-infected naïve animals were sequenced. Relative expression of relevant gene
products from both libraries was confirmed by reverse transcription real time PCR. Genes related to shrimp
immune response, such as the antimicrobial peptide penaeidin, crustin, C-type lectin, protease inhibitor and
chitin binding domain-containing protein were found in the library of pre-challenged shrimp. On the other
hand, genes from the naïve library coded for viral structural and functional proteins. To our knowledge, this is
the first report demonstrating that penaeidin-3 isoforms and crustin are over expressed in haemocytes of
WSSV-infected pre-challenged P. vannamei.
© 2009 Elsevier B.V. All rights reserved.
White Spot Syndrome Virus (WSSV) is the most devastating viral disease affecting cultivated shrimp aroundthe world. The CENIACUA breeding programme included host resistance as strategy to control WSSV. Since1999, all families were subjected to WSSV challenge tests and up to 5% of the animals survived. We suspectthat a unique genetic profile could be responsible for survival. By Subtractive Suppression Hybridizationimproved with Mirror Orientation Selection we compared gene expression in haemocytes of WSSV-infectedshrimp; seven were previously challenged (pre-challenged) and 10 naïve. Out of 1167 clones, 993 (85%) wereobtained from the WSSV-infected pre-challenged library, while 174 (15%) were from the WSSV-infected naïvelibrary. Differential expression was confirmed by hybridization using PCR DIG-labelled probes fromunsubtracted cDNA. The 216 (18.5%) clones differentially expressed in the WSSV-infected pre-challengedlibrary and 23 (2%) in the WSSV-infected naïve animals were sequenced. Relative expression of relevant geneproducts from both libraries was confirmed by reverse transcription real time PCR. Genes related to shrimpimmune response, such as the antimicrobial peptide penaeidin, crustin, C-type lectin, protease inhibitor andchitin binding domain-containing protein were found in the library of pre-challenged shrimp. On the otherhand, genes from the naïve library coded for viral structural and functional proteins. To our knowledge, this isthe first report demonstrating that penaeidin-3 isoforms and crustin are over expressed in haemocytes ofWSSV-infected pre-challenged P. vannamei.© 2009 Elsevier B.V. All rights reserved.
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