Identification of lysogenic phages in the native strain of bacillus subtilis

Shakiba Darvish alipour astaneh,1,* Morteza miri,2 Shamsozoha abolmaali,3 Nasrin shojaie,4

1. Semnan University
2. Dep. of Biotechnology, Semnan University, Semnan, Iran
3. Dep. of Biology, Faculty of Basic Science, Semnan University, Semnan, Iran
4. Dep. of Biotechnology, Semnan University, Semnan, Iran

Abstract


Introduction

Bacteriophages, lysogenic, show a high potential for the production of peptidoglycan hydrolyse peptides which lyse and destruct the cell wall of bacteria. therefore, they are effective tools in the treatment of bacterial infections.

Methods

To obtain the lysogenic(s) from ddbcc47; a native bacillus subtilis from semnan soil, six primer pairs were designed based on the genome of siphoviridea family- lysogenic for bacillus subtilis - using virus-host db database. the genome of bacteriophages was amplified with the primer pairs. the phages with positive amplicons were selected for further study. on the other hand, the antibacterial activity of ammonium sulfate purified endolysin obtained from ddbcc47 was measured by agar diffusion method.

Results

Based on our bioinformatics studies, the phage's blastoid, phage glittering, phage phi3t, phage riggi and phage vbbhas171 were distinguished as lysogens with no endolysin production. bacillus phage phi105, bacillus phage spbc2, bacteriophage spp1, and bacillus phage pm1, lysogenic in bacillus subtilis were found to produce endolysin. the results of the pcr reactions showed that the bacillus subtilis ddbcc47 was lysed by bacteriophage spp1 with the product of 816 bp endolysin gene.

Conclusion

Based on our bioinformatics studies, the phage's blastoid, phage glittering, phage phi3t, phage riggi and phage vbbhas171 were distinguished as lysogens with no endolysin production. bacillus phage phi105, bacillus phage spbc2, bacteriophage spp1, and bacillus phage pm1, lysogenic in bacillus subtilis were found to produce endolysin. the results of the pcr reactions showed that the bacillus subtilis ddbcc47 was lysed by bacteriophage spp1 with the product of 816 bp endolysin gene.

Keywords

Bacillus subtilis, lysogenic phages, endolysin, n-acetylmuramic-l-alanine amidase, siphoviridea