Creat A New Multi Locus Sequence Analysis Scheme (Mlsa) In Distinguishing The Two Bacteria Bacillus Subtilis And Bacillus Amyloliquefaciens And Compare To The 16s Ribosomal Rna Method

Abstract

A Multi Locus sequence analysis (MLSA) approach was established and performed on the two species that belong to Bacillus genus: Bacillus subtilis and Bacillus amyloliquefaciens, consisting of five strains (3 strains from B. subtilis and 2 strains from B. amyloliquefaciens) were provided by Biotechnology Center of Ho Chi Minh City. The study was based on sequences of 16S rRNA genes and the concatenate of five protein-coding housekeeping genes: glycerol uptake facilitator (glpF), phosphotransacetylase (ptA), phosphoribosyl aminoimidazole carboxamide formyl transferase (purH), pyruvate carboxylase (pycA), and RNA polymerase sigma factor (rpoD). After PCR amplification and sequencing, phylogenetic tree of 16S rRNA sequences and concatenate sequences (as well as phylogenetic tree of each housekeeping gene for supplementary) are constructed for comparison and discussion. The results show that using Multi Locus sequence analysis scheme can distinguish B. subtilis and B. amyloliquefaciens from each other and from other Bacillus species better than using traditional 16S rRNA genes. In conclusion, using Multi Locus sequence analysis scheme can reach for better resolution and differentiation of strains and can be developed in applying to the whole genus Bacillus.