Design of Specific Primer for Methallothionein Gene of Tor Fish (Tor tambra)

Abstract

The use of heavy metal biomarkers is needed to detect and integrate heavy metals at the molecular level. Metallothionein gene is a gene that is expressed in fish organs that have been heavily polluted. The aim of the study was to find specific primers for methallothionein of tor fish. Detection of the metallothionein gene expression in PCR instruments requires a primer in the form of a short chain DNA sequence as a specific target DNA identifier. The primer design was performed in silico using the NCBI site and multiple-aligned using Geneious Prime bioinformatic software. Primers were designed according to the conserved region of these genes. The primers specificity was checked using Primer BLAST tools in NCBI. The results showed that the forward primer 1 (5'- GAT TGC GCC AAG ACT GGA ACT –3') and reverse primer 1 (5' – ATC ACG TTG ACC TCC TCA CTG -3') qualified as good primers with an amplicon size 186 bp.