Molecular techniques for gene characterization: iPCR for identification of the 5'UTR of the ODC gene in the red macroalga Grateloupia imbricata

Abstract

iPCR or inverse Polymerase Chain Reaction is an amplification technique which allows in vitro amplification of DNA sequences flanking a known DNA region. This method is a variant of the common PCR method and it has been proved to become an important powerful tool in diverse research fields, such as genetic and molecular biology. In this study, iPCR technique was developed to investigate the 5' UTR (Untranslated Region) of the omithine decarboxylase (ODC, EC 4.1.1.17) gene in Grateloupia imbricata. As UTRs are involved in regulation, expression and transcriptional control of genes, the characterization and identification of this region represents a better approach to the gene 'machinery', which is still unclear. By performing this technique, a 242 bp fragment upstream from the gene-coding region was identified and sequenced. Alignment with Clustal Omega software finally confirmed it as a part of the flanking ODC 5 'UTR.