PCR Amplification, Sequencing of 16S rRNA Genes with Universal Primers and Phylogenetic Analysis of Pseudomonas aeruginosa

Pseudomonas aeruginosa is a gram negative bacterium isolated from soil. The isolate was cultured in a selective medium (Pseudomonas Agar base) at 37oC for 24 hours. For species identification, Pseudomonas like organisms creates lot of problems when identified with the help of morphological and biochemical characters. However, sequencing of 16S rRNA region is a suitable technique for species identification. The amplified product of 16S rRNA was submitted to NCBI database. Amplification of 16S rRNA gene region, and new sets of primer pairs were designed by NCBI database search tool. To study phylogenetic relationship between various strains of Pseudomonas aeruginosa have often been based on sequencing of 16S rRNA gene region. Distance tree was constructed to find out genetic similarity between the organisms. Hence gene sequencing of 16S rRNA region was a suitable technique to identify Pseudomonas aeruginosa at molecular level.