A metallothionein gene from hard clam Meretrix meretrix: Sequence features, expression patterns, and metal tolerance activities

Abstract

Metallothioneins (MTs) are low-molecular weight cytoplasmic heavy metal binding proteins. MTs can regulate the concentration of essential or non-essential metals in organisms, and have many important biological functions, including detoxification, trace element metabolism, and anti-oxidation. In the present study, we cloned and characterized a metallothionein gene (designated as MmMT) from the hard clam Meretrix meretrix. The complete cDNA sequence of MmMT contained an open reading frame (ORF) of 629 bp, which encoded a protein of 76 amino acids with a predicted molecular mass of 7.66 kDa and a calculated theoretical isoelectric point of 7.24. MmMT is highly similar to previously identified MTs from other species, with typical metallothionein features such as a high cysteine residue content and the absence of histidine and aromatic residues. The mRNA transcripts of MmMT were prevalent in all the tested tissues, and the expression levels of MmMT were highest in the hepatopancreas and hemocytes. During the stimulation of Vibrio splendidus, the mRNA transcripts of MmMT in the hepatopancreas and hemocytes were significantly increased. The Escherichia coli overexpressing MmMT performed strong growth in the media supplemented with CdCl2 and CuSO4 compared to the control strains. These results provide useful information for further investigation of the functions of MmMT in metal detoxification and the innate immune system.