Cloning of a calcitonin gene-related peptide from genomic DNA and its mRNA expression in flounder, Paralichthys olivaceus.
A part of genomic DNA including the calcitonin gene-related peptide (CGRP) gene was cloned from flounder by the genome-walking method. The intron/exon boundary was predicted to occur exactly at the same position as in salmon. The 37-amino acid molecule coded by the region from the intron/exon boundary to the stop codon was preceded by a typical Lys-Arg cleavage signal and included a cleavage/amidation site common to the CGRP of other vertebrates. The predicted amino acid sequence of flounder CGRP had 78%, 78%, 78%, 81%, and 73-78% identity to that of salmon, cod, frog, chicken, and mammalian CGRPs, respectively. Among vertebrates, CGRP is more conserved than calcitonin (CT) because the identity of flounder CT to mammalian CTs is 31-50%. Expression analysis indicated that this hormone is synthesized in the brain, heart, intestine, testis, and ovary. Since we have previously shown that the CGRP receptor is expressed in these tissues, it is suggested that CGRP secreted from each tissue functions in a paracrine or autocrine manner.
Suzuki
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N
Noto Marine Laboratory, Faculty of Science, Kanazawa University, Uchiura, 927-0553, Ishikawa, Japan. nobuo@kenroku.kanazawa-u.ac.jp
Suzuki
T
T
Kurokawa
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T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015740:Calcitonin Gene-Related Peptide / Q000097:blood / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D016384:Consensus Sequence; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005091:Exons; D005432:Flounder / Q000196:embryology / Q000235:genetics*; D015870:Gene Expression; D007438:Introns; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012474:Salmon / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014018:Tissue Distribution
D018076:DNA, Complementary; D012333:RNA, Messenger; D015740:Calcitonin Gene-Related Peptide