title: Molecular cloning and expression analysis of a thioredoxin from rock bream, Oplegnathus fasciatus, and biological activity of the recombinant protein.
abstract: Thioredoxins (TRxs) are a family of small, highly conserved proteins that are essential for the maintenance of cellular homeostasis. TRx1, which contains a conserved redox-active site, Cys-Gly-Pro-Cys, is a proinflammatory cytokine, B cell growth factor, macrophage migration inhibiting factor (MIF), and an immune regulatory cytokine. The TRx1 homologue cDNA was isolated from the rock bream LPS-stimulated liver cDNA library, RbTRx1. RbTRx1 consists of 730 bp full-length cDNA with a 324 bp open reading frame encoding 108 amino acids. When compared with other known TRx1 peptide sequences, the most conserved region of the RbTRx1 peptide was the redox-active site Cys-Gly-Pro-Cys. Phylogenetic analysis grouped the RbTRx1 with other vertebrate TRx1 peptides. Quantitative real-time PCR analysis revealed the presence of RbTRx1 transcripts in liver, gill, kidney, and muscle. The expression of RbTRx1 mRNA in kidney leukocytes was upregulated after bacterial and viral challenge. The kidney leukocytes were treated with a high concentration of rRbTRx1, which significantly enhanced cell proliferation (1 μg/ml and 10 μg/ml) and viability under oxidative stress (10 μg/ml).
authors.affiliation: Department of Aqualife Medicine, Chonnam National University, Yeosu, Republic of Korea; Fish Health Center, Chonnam National University, Yeosu, Republic of Korea.
authors.forename: Do-Hyung
authors.initials: DH
authors.lastname: Kim
authors.forename: Joo-Won
authors.initials: JW
authors.lastname: Kim
authors.forename: Ji-Min
authors.initials: JM
authors.lastname: Jeong
authors.forename: Hyung-Jun
authors.initials: HJ
authors.lastname: Park
authors.forename: Chan-Il
authors.initials: CI
authors.lastname: Park
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004266:DNA Virus Infections / Q000276:immunology / Q000662:veterinary; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D020609:Edwardsiella tarda / Q000502:physiology; D004756:Enterobacteriaceae Infections / Q000276:immunology / Q000662:veterinary; D004926:Escherichia coli / Q000235:genetics; D005393:Fish Diseases / Q000276:immunology*; D029941:Fish Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D020869:Gene Expression Profiling; D015723:Gene Library; D017994:Iridovirus / Q000502:physiology; D007668:Kidney / Q000276:immunology / Q000378:metabolism; D008099:Liver / Q000276:immunology / Q000378:metabolism; D008969:Molecular Sequence Data; D010473:Perciformes / Q000235:genetics* / Q000276:immunology / Q000378:metabolism* / Q000382:microbiology; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013290:Streptococcal Infections / Q000276:immunology / Q000662:veterinary; D013291:Streptococcus / Q000502:physiology; D013879:Thioredoxins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D029941:Fish Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D013879:Thioredoxins