Isolation of Antimicrobial Genes from Oryza rufipogon Griff by Using a Bacillus subtilis Expression System with Potential Antimicrobial Activities.

International Journal of Molecular Sciences
Jiale LiWubei Dong

Abstract

Antimicrobial genes are distributed in all forms of life and provide a primary defensive shield due to their unique broad-spectrum resistance activities. To better isolate these genes, we used the Bacillus subtilis expression system as the host cells to build Oryza rufipogon Griff cDNA libraries and screen potential candidate genes from the library at higher flux using built-in indicator bacteria. We observed that the antimicrobial peptides OrR214 and OrR935 have strong antimicrobial activity against a variety of Gram-positive and Gram-negative bacteria, as well as several fungal pathogens. Owing to their high thermal and enzymatic stabilities, these two peptides can also be used as field biocontrol agents. Furthermore, we also found that the peptide OrR214 (MIC 7.7-10.7 μM) can strongly inhibit bacterial growth compared to polymyxin B (MIC 5-25 μM) and OrR935 (MIC 33-44 μM). The cell flow analysis, reactive oxygen burst, and electron microscopy (scanning and transmission electron microscopy) observations showed that the cell membranes were targeted by peptides OrR214 and OrR935, which revealed the mode of action of bacteriostasis. Moreover, the hemolytic activity, toxicity, and salt sensitivity experiments demonstrated that th...Continue Reading

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Citations

Apr 4, 2021·International Journal of Molecular Sciences·Mohamed F HassanWubei Dong

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Methods Mentioned

BETA
Assay
scanning
transmission electron microscopy
confocal
electron microscopy
PCR
ELISA
fluorescence
confocal microscopy

Software Mentioned

FlowJo
BLAST

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