Molecular cloning and functional characterization of a mouse ccl6 analog gene in the rat

• Published in: Genetics and molecular research 2012-01, Vol.11 (4), p.3889-3898
• Main Authors: Jiang, N, Zheng, Y H, Chen, X J, Qiu, C, Zhang, X F, Wen, S H, Bian, G X
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Chemokines, CC - chemistry; Chemokines, CC - genetics; Chemokines, CC - isolation & purification; Chemotaxis - drug effects; Cloning, Molecular; Electrophoresis, Polyacrylamide Gel; Exons - genetics; Hybridization, Genetic - drug effects; Introns - genetics; Mice; Molecular Sequence Data; Neutrophils - cytology; Neutrophils - drug effects; Phylogeny; Rats; Recombinant Proteins - pharmacology; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Nucleic Acid
• ISSN: 1676-5680; 1676-5680
• DOI: 10.4238/2012.November.12.6

Suppression subtractive hybridization was used to analyze differential expression of genes in rat peritoneal macrophages after granulocyte macrophage colony-stimulating factor treatment. We identified and cloned the mouse C10 analog gene in the rat, and named it as ccl6. The full-length cDNA of rat ccl6 was 467 bp, which contains a single-open reading frame and encodes 116 amino acid residues. Compared with other C-C chemokines, the rat ccl6 gene had an unusual four-exon genome structure instead of the typical three exons, it had the highest homology with murine ccl6. The rat ccl6 gene was localized on chromosome 10, where most of the C-C chemokine superfamily members are located. The recombinant rat C-C chemokine ligand 6 (CCL6) protein was expressed by the pGEX4T-1 plasmid in Escherichia coli BL21. The purified recombinant protein had bioactivity similar to that of mouse CCL6, which is a chemoattractant for macrophages and lymphocytes, but not for neutrophils.