NDLI: Identification of Bacillus Probiotics Isolated from Soil Rhizosphere Using 16S rRNA, recA, rpoB Gene Sequencing and RAPD-PCR

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Identification of Bacillus Probiotics Isolated from Soil Rhizosphere Using 16S rRNA, recA, rpoB Gene Sequencing and RAPD-PCR

Content Provider	Springer Nature Link
Author	Mohkam, Milad Nezafat, Navid Berenjian, Aydin Mobasher, Mohammad Ali Ghasemi, Younes
Copyright Year	2016
Abstract	Some Bacillus species, especially Bacillus subtilis and Bacillus pumilus groups, have highly similar 16S rRNA gene sequences, which are hard to identify based on 16S rDNA sequence analysis. To conquer this drawback, rpoB, recA sequence analysis along with randomly amplified polymorphic (RAPD) fingerprinting was examined as an alternative method for differentiating Bacillus species. The 16S rRNA, rpoB and recA genes were amplified via a polymerase chain reaction using their specific primers. The resulted PCR amplicons were sequenced, and phylogenetic analysis was employed by MEGA 6 software. Identification based on 16S rRNA gene sequencing was underpinned by rpoB and recA gene sequencing as well as RAPD-PCR technique. Subsequently, concatenation and phylogenetic analysis showed that extent of diversity and similarity were better obtained by rpoB and recA primers, which are also reinforced by RAPD-PCR methods. However, in one case, these approaches failed to identify one isolate, which in combination with the phenotypical method offsets this issue. Overall, RAPD fingerprinting, rpoB and recA along with concatenated genes sequence analysis discriminated closely related Bacillus species, which highlights the significance of the multigenic method in more precisely distinguishing Bacillus strains. This research emphasizes the benefit of RAPD fingerprinting, rpoB and recA sequence analysis superior to 16S rRNA gene sequence analysis for suitable and effective identification of Bacillus species as recommended for probiotic products.
Starting Page	8
Ending Page	18
Page Count	11
File Format	PDF
ISSN	18671306
Journal	Probiotics and Antimicrobial Proteins
Volume Number	8
Issue Number	1
e-ISSN	18671314
Language	English
Publisher	Springer US
Publisher Date	2016-02-22
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Bacillus Concatenation Probiotics Multilocus sequence analysis (MLSA) rpoB recA Randomly amplified polymorphic DNA-PCR (RAPD-PCR) Chemistry/Food Science Applied Microbiology Microbiology Protein Science Nutrition
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Molecular Medicine Microbiology

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