Molecular cloning, characterization and expression analysis of a CC chemokine gene from miiuy croaker (Miichthys miiuy)

Chemokines are a family of structurally related chemotactic cytokines that regulate the migration of leukocytes, under both physiological and inflammatory conditions. A partial cDNA of CC chemokine gene designed as Mimi-CC3 was isolated from miiuy croaker ( Miichthys miiuy ) spleen cDNA library. Unk...

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Bibliographic Details
Published in:Fish physiology and biochemistry 2012-12, Vol.38 (6), p.1697-1708
Main Authors: Cheng, Yuanzhi, Sun, Yuena, Shi, Ge, Wang, Rixin, Xu, Tianjun
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Animal Anatomy
Animal Biochemistry
Animal Physiology
Animals
Base Sequence
Biomedical and Life Sciences
Chemokines
Chemokines, CC - genetics
Chemokines, CC - metabolism
Cloning
Cloning, Molecular
Cytokines
Fish
Freshwater & Marine Ecology
Gene Expression Regulation - physiology
Genes
Genomics
Histology
Immune system
Life Sciences
Marine
Miichthys miiuy
Molecular Sequence Data
Morphology
Pathogens
Perciformes - metabolism
Phylogenetics
Phylogeny
Real-Time Polymerase Chain Reaction
RNA, Messenger - genetics
RNA, Messenger - metabolism
Spleen
Vibrio - physiology
Vibrio anguillarum
Vibrio Infections - immunology
Vibrio Infections - metabolism
Vibrio Infections - veterinary
Zoology
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Summary:Chemokines are a family of structurally related chemotactic cytokines that regulate the migration of leukocytes, under both physiological and inflammatory conditions. A partial cDNA of CC chemokine gene designed as Mimi-CC3 was isolated from miiuy croaker ( Miichthys miiuy ) spleen cDNA library. Unknown 3′ part of the cDNA was amplified by 3′-RACE. The complete cDNA of Mimi-CC3 contains an 89-nt 5′-UTR, a 303-nt open reading frame and a 441-nt 3′-UTR. Three exons and two introns were identified in Mimi-CC3. The deduced Mimi-CC3 protein sequences contain a 22 amino acids signal peptide and a 78 amino acids mature polypeptide, which possesses the typical arrangement of four cysteines as found in other known CC chemokines. It shares low amino acid sequence identities with most other fish and mammalian CC chemokines (less than 54.1 %), but shares very high identities with large yellow croaker CC chemokine (94.6 %). Phylogenetic analysis showed that Mimi-CC3 gene may have an orthologous relationship with mammalian/amphibian CCL25 gene. Tissue expression distributed analysis showed that Mimi-CC3 gene was constitutively expressed in all nine tissues examined, although at different levels. Upon stimulated with Vibrio anguillarum , the time-course analysis using a real-time PCR showed that Mimi-CC3 transcript in kidney and liver was obviously up-regulated and reached the peak levels, followed by a recovery. Mimi-CC3 expression in kidney was more strongly increased than in liver. However, down-regulation was observed in spleen. These results indicated that Mimi-CC3 plays important roles in miiuy croaker immune response as well as in homeostatic mechanisms.
ISSN:0920-1742
1573-5168
DOI:10.1007/s10695-012-9665-6