Epigenetic Segregation of Microbial Genomes from Complex Samples Using Restriction Endonucleases HpaII and McrB
| Autor: | Helen E. Barnes, Paula King, Long K. Pham, Guohong Liu, R. Allyn Forsyth, Robert T. Yamamoto, Dan Sphar, Christopher Q. Weston, Shannon Waltz |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
DNA Bacterial Genetics Microbial DNA Plant HpaII 030106 microbiology lcsh:Medicine Genomics Biology Deoxyribonuclease HpaII Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Humans Genomic library DNA Fungal lcsh:Science Gene Library Genetics Multidisciplinary Escherichia coli Proteins Microbiota lcsh:R Sputum DNA Restriction Enzymes Sequence Analysis DNA DNA Methylation DNA Protozoan Restriction enzyme 5-Methylcytosine 030104 developmental biology chemistry Metagenomics DNA methylation DNA Viral Metagenome CpG Islands lcsh:Q Research Article |
| Zdroj: | PLoS ONE, Vol 11, Iss 1, p e0146064 (2016) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | We describe continuing work to develop restriction endonucleases as tools to enrich targeted genomes of interest from diverse populations. Two approaches were developed in parallel to segregate genomic DNA based on cytosine methylation. First, the methyl-sensitive endonuclease HpaII was used to bind non-CG methylated DNA. Second, a truncated fragment of McrB was used to bind CpG methylated DNA. Enrichment levels of microbial genomes can exceed 100-fold with HpaII allowing improved genomic detection and coverage of otherwise trace microbial genomes from sputum. Additionally, we observe interesting enrichment results that correlate with the methylation states not only of bacteria, but of fungi, viruses, a protist and plants. The methods presented here offer promise for testing biological samples for pathogens and global analysis of population methylomes. |
| Databáze: | OpenAIRE |
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