title: A manganese superoxide dismutase in blood clam Tegillarca granosa: molecular cloning, tissue distribution and expression analysis.
abstract: Superoxide dismutase (SOD, EC 1.15.1.1) is one of the central enzymes involved in scavenging the high level of reactive oxygen species (ROS) by transforming O₂⁻ into hydrogen peroxide and oxygen. The full-length mitochondrial Mn-SOD cDNA of blood clam Tegillarca granosa (denoted as TgmMnSOD) was identified from haemocytes by homology cloning and rapid amplification of cDNA ends (RACE) approaches. The nucleotide sequence of TgmMnSOD consisted of 1106bp with a 5' UTR of 195bp, a 3' UTR of 227bp with a candidate polyadenylation signal sequence ATTAAA and a short polyA tail, and an open reading frame (ORF) of 648bp encoding a secreted polypeptide of 227 amino acids residues. The deduced amino acid sequence of TgmMnSOD shared significant homology to mMnSODs from other species, indicating that TgmMnSOD should be a novel member of the mMnSOD family. Several highly conserved motifs including three mMnSOD signatures, amino acid residues responsible for coordinating the manganese and the putative active center were almost completely conserved in the deduced amino acid of TgmMnSOD. The mRNA expression of TgmMnSOD in the tissues of mantle, foot, gill, haemocytes and hepatopancreas was examined by quantitative real-time PCR (qT-PCR) and mRNA transcripts of TgmMnSOD were mainly detected in hepatopancreas, haemocytes, and gill and weakly detected in the mantle and foot. The temporal expression of TgmMnSOD in haemocytes after heavy metal exposure revealed that TgmMnSOD could be induced by the three pollutants with different response profiles. The polyclonal antibodies generated from the recombinant product of TgmMnSOD could specifically identify not only the recombinant product, but also the native protein from haemocytes. The present results strongly suggested that TgmMnSOD was a cute response protein involved in marine heavy metal contaminants challenge in T. granosa.
authors.affiliation: Faculty of Life Science and Biotechnology, Ningbo University, Fenghua Road, Ningbo, Zhejiang Province, PR China.
authors.forename: Chenghua
authors.initials: C
authors.lastname: Li
authors.forename: Jingjing
authors.initials: J
authors.lastname: He
authors.forename: Xiurong
authors.initials: X
authors.lastname: Su
authors.forename: Taiwu
authors.initials: T
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D049872:Bivalvia / Q000201:enzymology* / Q000235:genetics; D015153:Blotting, Western; D003001:Cloning, Molecular; D015870:Gene Expression; D006434:Hemocytes / Q000187:drug effects / Q000201:enzymology; D019216:Metals, Heavy / Q000633:toxicity; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014018:Tissue Distribution
chemical_list: D019216:Metals, Heavy; D012333:RNA, Messenger; D013482:Superoxide Dismutase