Article ID Journal Published Year Pages File Type
2429227 Developmental & Comparative Immunology 2013 8 Pages PDF
Abstract

•RNA interference and Suppression Subtractive Hybridization techniques were combined to identify functional genes in shrimp.•Genes regulated by Relish were isolated after screening of the SSH library.•The selected candidates were verified by qPCR.•The present data provides us a wide view to understand the function of Relish gene in the innate immunity of shrimp.

Relish is a key NF-κB transcription factor in the innate immunity. Learning the function of Relish in regulating the related genes of shrimp will be helpful to understand the shrimp immunity. In the present study, RNA interference (RNAi) and suppression subtractive hybridization (SSH) techniques were combined together to identify the genes regulated by Relish. A forward SSH library represents the genes whose transcription was regulated by Relish, and the reverse SSH library represents the genes whose transcription was up-regulated after Relish was silenced in shrimp responsive to Vibrio anguillarium (VA) stimulation. In the forward library, 43 unique genes were identified, and in the reverse library, 57 genes were identified. The expression of ten differentially expressed genes, including early cuticle protein5 (ECP5), Toll-like receptor protein (TLRP), antiviral factor (AV), C-type lectin receptor (CLR), thrombospondin (TSP), S-adenosylmethionine synthetase (SAMS), carcinolectin 5b-5 (CL5b-5), QM protein (QMP), heat shock protein 67B2 (HSP67B2), and Thioredoxin-related protein 14 (TRP14) were further confirmed by real-time PCR. The present data provides us a wide view to understand the function of Relish gene in the innate immunity of shrimp.

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