- Volume 5 Issue 2
Molecular cloning and characterization of metallothionein cDNA gene in channel catfish
챠넬메기의 metallothionein cDNA 유전자의 cloning 및 그 특성에 관한 연구
- Lee, In-Jung (Department of Microbiology, National Fisheries University of Pusan) ;
- Song, Young-Hwan (Department of Microbiology, National Fisheries University of Pusan)
- Published : 1992.12.30
Metallothionein is an essential and common protein to regulate the intracellular concentration of heavy metals, which exist in most organisms from bacteria to vertebrates. Although the detailed function of metallothianein has not been fully identified until yet, it may be involoved in the cellular protection against the heavy metal toxicity and in the global regulation of several other genes and the expression of metalloproteins. We have cloned the full cDNA clone of metallothionein gene in Channel Catfish by Reverse Transcriptase-Polymerase Chain Reaction(RT-PCR) starting from poly(A)-containing mRNAs. All PCR fragments have been subcloned into EcoRV site of pBluescript SK+ and dT-tailed at Smal site of pUC19, then PCR products are recovered by the double digestion of recombinant plasmids wiht EcoRI and HindIII, which are adjacent to EcoRV site in multicloning sites or by rapid PCR screening. The nucleotide sequence analysis of pMT150(one of the PCR clones) showed high homology with several other piscine metallothionein cDNA genes.