High-Quality Nucleic Acid Isolation from Hard-to-Lyse Bacterial Strains Using PMAP-36, a Broad-Spectrum Antimicrobial Peptide
| Autor: | Munjeong Choi, Hyoim Jeon, Hye-sun Cho, Kwonho Hong, Jin-Hoi Kim, Chankyu Park, Yunjung Lee, Nagasundarapandian Soundrarajan, Byeongyong Ahn |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
DNA Bacterial Staphylococcus aureus Lysis QH301-705.5 Antimicrobial peptides Cell Fractionation Catalysis Article Microbiology Inorganic Chemistry 03 medical and health sciences antimicrobial peptides 0302 clinical medicine PMAP-36 Physical and Theoretical Chemistry Biology (General) Molecular Biology QD1-999 Spectroscopy nucleic acids isolation biology Chemistry Organic Chemistry RNA General Medicine Antimicrobial biology.organism_classification Computer Science Applications genomic DNA RNA Bacterial 030104 developmental biology 030220 oncology & carcinogenesis Nucleic acid RNA extraction Bacteria Antimicrobial Cationic Peptides |
| Zdroj: | International Journal of Molecular Sciences International Journal of Molecular Sciences, Vol 22, Iss 4149, p 4149 (2021) Volume 22 Issue 8 |
| ISSN: | 1422-0067 |
| Popis: | The efficiency of existing cell lysis methods to isolate nucleic acids from diverse bacteria varies depending on cell wall structures. This study tested a novel idea of using broad-spectrum antimicrobial peptides to improve the lytic efficiency of hard-to-lyse bacteria and characterized their differences. The lysis conditions of Staphylococcus aureus using recombinant porcine myeloid antimicrobial peptide 36 (PMAP-36), a broad-spectrum pig cathelicidin, was optimized, and RNA isolation was performed with cultured pellets of ten bacterial species using various membranolytic proteins. Additionally, three other antimicrobial peptides, protegrin-1 (PG-1), melittin, and nisin, were evaluated for their suitability as the membranolytic agents of bacteria. However, PMAP-36 use resulted in the most successful outcomes in RNA isolation from diverse bacterial species. The amount of total RNA obtained using PMAP-36 increased by ~2-fold compared to lysozyme in Salmonella typhimurium. Streptococci species were refractory to all lytic proteins tested, although the RNA yield from PMAP-36 treatment was slightly higher than that from other methods. PMAP-36 use produced high-quality RNA, and reverse transcription PCR showed the efficient amplification of the 16S rRNA gene from all tested strains. Additionally, the results of genomic DNA isolation were similar to those of RNA isolation. Thus, our findings present an additional option for high quality and unbiased nucleic acid isolation from microbiomes or challenging bacterial strains. |
| Databáze: | OpenAIRE |
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