Molecular Cloning, Sequence Analysis and Prokaryotic Expression of Sweetfish Macrophage Inflammatory Protein-2 Gene

Abstract

Abstract: Macrophage inflammatory protein-2（MIP-2）is an important chemokine, which promotes neutrophils to be the sites of inflammation and eliminates the inflammatory response. In this study, the sequence of MIP-2 gene from de-novo transcriptome sequencing of sweetfish macrophages was obtained. An open reading frame was composed of 318 nucleotides, which encoded a protein of 105 amino acids with a molecular weight of 11.6 kD. Its N-terminal 19 residues were the signal peptides. Sequence alignment showed that sweetfish MIP-2 shared the highest homology with the MIP-2 from Esox lucius with 54% protein sequence identity. In healthy sweetfish, MIP-2 mRNA was mainly expressed in the spleen, kidney, brain and gills, weakly expressed in the liver, heart, muscle and intestine. Quantitative RT-PCR analysis showed that MIP-2 transcripts were significantly up-regulated in the tested tissues, especially the kidney and muscle. The results suggested that MIP-2 might play an important role in sweetfish antibacterial immunity.

Key words: MIP-2, Sweetfish, Listonella anguillarum infection, Gene expression

Wang Shihui, Shi Yuhong, Chen Jiong. Molecular Cloning, Sequence Analysis and Prokaryotic Expression of Sweetfish Macrophage Inflammatory Protein-2 Gene[J]. Biotechnology Bulletin, 2014, 0(7): 143-149.

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