Phylogenetic tree of Proteus spp. Based on partial rpoB gene sequence analysis

Abstract

Due to the importance of the proposed rpoB gene as an alternative biomarker for microbial community studies, this study has focused on phylogenetic relationships among local Proteus clinical isolates. Fifty bacterial isolates were collected and identified phenotypically according to the culture, microscopic examination and biochemical tests. VITEK 2 compact system was used to confirm identification. Genotypic identification was performed after DNA extraction for 10 selected isolates and amplification with rpoB gene-specific primer and gel electrophoresis. The products were detected with a ( 1090 bp ) molecular size band, which was sent for Sanger sequencing using an ABI3730XL automated DNA sequencer, and data were analyzed and compared with standard sequences in GenBank.The isolates have been registered in the National Center for Biotechnology Information (NCBI) with accession numbers and named (HIMUS1, HIMUS2, HIMUS3, HIMUS4, HIUS5, HIMUS6, HIMUS7, HIMUS8, HIMUS9 and HIMUS10 ). The phylogenetic tree was constructed using partial (895 bp) rpoB gene sequences for those ten strains. Evolutionary distances were calculated using the method of Maximum Composite Likelihood with 1000 bootstrap replicates using GENEIOUS software. The sequences presented a similarity percentage ranging between (98.76% and 100%) when compared with the sequences of standard strains in NCBI. Keywords: rpoB gene, Proteus spp. Sequencing, Phylogenetic analysis