Cloning of Dehd Gene Into an Expression Vector Puc19 for Potential Application in Bioremediation

Dehalogenase D (dehD) is produced by Rhizobium sp. RC1 which breakdown carbon-halogen bond of Dhalogenated compounds which are known recalcitrant chemicals. The present research intends to clone dehD into a suitable expression vector pUC19 for possible use in bioremediation of recalcitrant chemicals such as Dhalogenated compounds. The dehD gene of Rhizobium sp. RC1 sequence was retrieved from GenBank, synthesized by Genscript, USA. The gene construct in pUC57 and pUC19 plasmid were digested using BamHI and HindIII restriction enzymes and ligated using Quick T4 DNA ligase which allows ligation of DNA fragments with blunt end or cohesive end within 5 min at 25 OC. The clone was re-digested using BamHI and HindIII and analyzed using agarose gel electrophoresis and sequencing technique. It was established that dehD was cloned into E. coli using pUC19 vector. This was necessitated in order to obtain dehalogenase enzyme with enhanced potentials in environmental bioremediation, for industrial production of chemicals, pharmaceutical and medical applications. This is the first report on cloning of dehD gene for possible application in bioremediation.