Cloning and sequencing of putative beta-lactam genes from amoeba (Acanthamoeba spp.)

Abstract

Acanthamoeba spp. has been known as sources for anti-microbe compounds. Antibiotic penicillin and cephalosporin under the group of beta-lactam antibiotics are important as antibacterial treatment for bacterial infection. Previous study had successfully identity twelve putative fragments with PCR technique. The putative fragments consist beta-lactam gene were cloned and sequenced. Two selected putative fragments, Spnl-1 (1500bp) and Spn5-1 (900bp) were re-amplified and cloned into the pGEM-T vector. The putative recombinant colonies were screened by using colony PCR technique. The positive recombinant colonies were selected for plasmid extraction and further confirm the presence of the inserted DNA with PCR technique. The complete nucleotide sequence of clone pSpnl-1 consists of 1494bp while the deduced amino acids from the complete nucleotide are 498 amino acids. The translated acid amino for clone pSpnl-1 shows 39 - 43% positive similarity and 23 28% identity to different protein sequence from different species in the Gene Bank database.