Molecular cloning and expression of preproadrenomedullin gene from common carp Cyprinus carpio L.
A preproadrenomedullin (preproAM) gene was isolated from common carp Cyprinus carpio L. by PCR mediated homology cloning. The isolated gene was composed of 516bp, which translated to a protein having 171 amino acid residues. The signal peptide was composed of 23 amino acid residues and predicted adrenomedullin (AM) was conserved in this sequence and made up of 49 amino acid residues. AM gene from carp shared identities of 64.0, 44.2, and 46.0% to takifugu (AM-1), human, and rat AM orthologues, respectively. The analysis of genomic structure revealed that carp preproAM gene is spread over two exons interrupted by one intron (92bp). AM genes clustered together and carp AM gene showed similarity to takifugu AM-1 gene according to the phylogenetic analysis. Expression analysis revealed that carp AM gene expressed constitutively in all the organs confirmed. Moreover, we demonstrated that expression of carp AM gene increased by treatment of LPS or hydrocortisone in different tissues by quantitative real-time PCR.
Kono
Tomoya
T
Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.
Sakai
Masahiro
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002347:Carps / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D006854:Hydrocortisone / Q000494:pharmacology; D007668:Kidney / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D011333:Pro-Opiomelanocortin / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution
D018076:DNA, Complementary; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D011333:Pro-Opiomelanocortin; D006854:Hydrocortisone