Cloning and sequencing of putative anhydrotetracycline oxygenase (Ahtm) gene from amoeba (Acanthamoeba spp.)

Abstract

Acanthamoeba spp. was believed to possess vanous antimicrobial peptides and proteins to combat bacterial growth inside their phagosomes. Tetracycline belongs to a group of antibiotics that contain antimicrobial and pharmacokinetic properties. Anhydrotetracycline oxygenase (anhydrotetracycline monooxygenase or AHTM) is the enzyme that involve in the reaction of biosynthesis pathway of Streptomyces spp. in the production of tetracycline. Previous studies had successfully identified six putative fragments by using PCR technique. The putative fragment of AHTM5 was selected and re-amplified and cloned into pGEM-T Easy Vector. The putative recombinant colonies were selected for plasmid extraction. The presence of inserted DNA was confirmed by using PCR technique. The complete nucleotide sequence consists of 243bp while the deduced amino acids from the complete nucleotide sequence are 81. The translated amino acid sequence of clone pAHTM5 shows 81 %85%