Cloning and sequencing of ABC transporter ATP-binding protein encoding gene from Streptomyces minoensis

Abstract

Bacterial products are secreted or transferred out of the cell via transporter systems such as ABC transporters. Nucleotide analysis and comparison of the Nucleotide Binding Domain (NBD) of the ABC transporter ATP-binding protein encoding gene from Streptomyces minoensis, using PCR method is described. Following the culture of bacterium and genomic DNA isolation, the required fragment of the DNA was amplified using PCR technique. The amplified DNA was then cloned into pTZ57R/T vector. E. coli: DH5? competent cells were transformed and the cloned fragment of DNA was then sequenced, resulting in identification and submission of a 913 nucleotide chain from ABC transporter ATP-binding protein encoding gene from Streptomyces minoensis in NCBI under accession number of DQ388679. The blast results of submitted sequence shows high homology in nucleotide and amino acid levels with ABC transporter ATP-binding protein encoding gene/protein from other Streptomyces species. Bioinformatics analysis revealed that this amino acid chain contains all of the functional motifs of NBD defined in other investigated ABC transporters ATP-binding proteins. é 2008 Asian Network for Scientific Information.