TY - JOUR
T1 - Thioredoxin of golden pompano involved in the immune response to Photobacterium damselae
AU - Wang, Long
AU - Guo, Huayang
AU - zhang, Nan
AU - Ma, Zhenhua
AU - Jiang, Shigui
AU - Zhang, Dianchang
PY - 2015/8
Y1 - 2015/8
N2 - Thioredoxin (TRX) is one of the key systems responsible for keeping the intracellular environment in a highly reduced state. In this study, a full-length TRX cDNA sequence (ToTRX) from golden pompano Trachinotus ovatus was identified after pyrosequencing of golden pompano cDNA library. ToTRX cDNA is comprised of 786bp, and contained a 324bp open reading frame (ORF) encoding a 107 amino acid polypeptide, a 5' untranslated region (UTR) of 116bp, and a long 3'- UTR of 346bp. Multiple sequence alignment revealed that ToTRX contained the highly conserved redox active disulphide/dithiol site (CGPC) of the thioredoxin active family, and phylogenetic tree showed that ToTRX had a closer evolution relationship with TRX from Oplegnathus fasciatus and Anoplopoma fimbria. ToTRX mRNA is ubiquitously expressed in all detected tissues with the higher expression levels in the stomach, gill and fin tissues. The expression of ToTRX mRNA was significantly up-regulated in liver, kidney, intestine and spleen of golden pompano injected with Photobacterium damselae. The recombinant ToTRX protein (rToTRX) was expressed in Escherichia coli BL21 (DE3), and then purified and refolded. The insulin disulfides assay was performed to investigate the enzymatic oxidoreductase activity of rToTRX, and the results demonstrated that rToTRX exhibited a high reducing activity in presence of DTT, while no activity was observed in the regroup without DTT and blank control group. Over all, the study provided the useful information to help further understand the functional mechanism of TRX in marine fish immunity.
AB - Thioredoxin (TRX) is one of the key systems responsible for keeping the intracellular environment in a highly reduced state. In this study, a full-length TRX cDNA sequence (ToTRX) from golden pompano Trachinotus ovatus was identified after pyrosequencing of golden pompano cDNA library. ToTRX cDNA is comprised of 786bp, and contained a 324bp open reading frame (ORF) encoding a 107 amino acid polypeptide, a 5' untranslated region (UTR) of 116bp, and a long 3'- UTR of 346bp. Multiple sequence alignment revealed that ToTRX contained the highly conserved redox active disulphide/dithiol site (CGPC) of the thioredoxin active family, and phylogenetic tree showed that ToTRX had a closer evolution relationship with TRX from Oplegnathus fasciatus and Anoplopoma fimbria. ToTRX mRNA is ubiquitously expressed in all detected tissues with the higher expression levels in the stomach, gill and fin tissues. The expression of ToTRX mRNA was significantly up-regulated in liver, kidney, intestine and spleen of golden pompano injected with Photobacterium damselae. The recombinant ToTRX protein (rToTRX) was expressed in Escherichia coli BL21 (DE3), and then purified and refolded. The insulin disulfides assay was performed to investigate the enzymatic oxidoreductase activity of rToTRX, and the results demonstrated that rToTRX exhibited a high reducing activity in presence of DTT, while no activity was observed in the regroup without DTT and blank control group. Over all, the study provided the useful information to help further understand the functional mechanism of TRX in marine fish immunity.
KW - Antioxidant function
KW - Immune response
KW - MRNA expression
KW - Thioredoxin
KW - Trachinotus ovatus
UR - http://www.scopus.com/inward/record.url?scp=84935001697&partnerID=8YFLogxK
U2 - 10.1016/j.fsi.2015.05.044
DO - 10.1016/j.fsi.2015.05.044
M3 - Article
C2 - 26052015
AN - SCOPUS:84935001697
SN - 1050-4648
VL - 45
SP - 808
EP - 816
JO - Fish and Shellfish Immunology
JF - Fish and Shellfish Immunology
IS - 2
ER -