Review
- MINIREVIEW] On the study of microbial transcriptomes using second- and third-generation sequencing technologies
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Sang Chul Choi
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J. Microbiol. 2016;54(8):527-536. Published online August 2, 2016
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DOI: https://doi.org/10.1007/s12275-016-6233-2
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Abstract
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Second-generation sequencing technologies transformed the
study of microbial transcriptomes. They helped reveal the
transcription start sites and antisense transcripts of microbial
species, improving the microbial genome annotation.
Quantification of genome-wide gene expression levels allowed
for functional studies of microbial research. Ever-evolving
sequencing technologies are reshaping approaches to studying
microbial transcriptomes. Recently, Oxford Nanopore
Technologies delivered a sequencing platform called MinION,
a third-generation sequencing technology, to the research
community. We expect it to be the next sequencing technology
that enables breakthroughs in life science fields. The studies
of microbial transcriptomes will be no exception. In this paper,
we review microbial transcriptomics studies using second-
generation sequencing technology. We also discuss the
prospect of microbial transcriptomics studies with thirdgeneration
sequencing.
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Journal Article
- Effect of pH on Conjugated Linoleic Acid (CLA) Formation of Linolenic Acid Biohydrogenation by Ruminal Microorganisms
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Yongjae Lee
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J. Microbiol. 2013;51(4):471-476. Published online August 30, 2013
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DOI: https://doi.org/10.1007/s12275-013-1070-z
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Abstract
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Conventional beliefs surrounding the linolenic acid (LNA; cis-9 cis-12 cis-15 C18:3) biohydrogenation (BH) pathway propose that it converts to stearic acid (SA) without the formation of conjugated linoleic acid (CLA) as intermediate isomers. However, an advanced study (Lee and Jenkins, 2011) verified that LNA BH yields multiple CLAs. This study utilized the stable isotope tracer to investigate the BH intermediates of 13C-LNA with different pH conditions (5.5 and 6.5). The 13C enrichment was calculated as a 13C/12C ratio of labeled minus unlabeled. After 24 h, eight CLA isomers were significantly enriched on both pH treatment, this result verifies that these CLAs originated from 13C-LNA BH which supports the results of Lee and Jenkins (2011). The enrichment of cis-cis double bond CLAs (cis-9 cis-11 and cis-10 cis-12 CLA) were significantly higher at low pH conditions. Furthermore, the concentration of cis-10 cis-12 CLA at low pH was four times higher than at high pH conditions after a 3 h incubation. These differences support the LNA BH pathways partial switch under different pH conditions, with a strong influence on the cis-cis CLA at low pH. Several mono-, di-, and tri-enoic fatty acid isomers were enriched during 24 h of incubation, but the enrichment was decreased or restricted at low pH treatment. Based on these results, it is proposed that low pH conditions may cause a changed or limited capacity of the isomerization and reduction steps in BH.
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- Invited review: Research on ruminal biohydrogenation—Achievements, gaps in knowledge, and future approaches from the perspective of dairy science
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Research Support, U.S. Gov't, Non-P.H.S.
- Identification of Enriched Conjugated Linoleic Acid Isomers in Cultures of Ruminal Microorganisms after Dosing with 1-13C-Linoleic Acid
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Yong-Jae Lee , Thomas C. Jenkins
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J. Microbiol. 2011;49(4):622-627. Published online September 2, 2011
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DOI: https://doi.org/10.1007/s12275-011-0415-8
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43
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9
Scopus
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Abstract
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Most studies of linoleic acid biohydrogenation propose that it converts to stearic acid through the production
of cis-9 trans-11 CLA and trans-11 C18:1. However, several other CLA have been identified in ruminal
contents, suggesting additional pathways may exist. To explore this possibility, this research investigated
the linoleic acid biohydrogenation pathway to identify CLA isomers in cultures of ruminal microorganisms
after dosing with a 13C stable isotope. The 13C enrichment was calculated as [(M+1/M)×100] in labeled
minus unlabeled cultures. After 48 h incubation, significant 13C enrichment was observed in seven CLA
isomers, indicating their formation from linoleic acid. All enriched CLA isomers had double bonds in either
the 9,11 or 10,12 position except for trans-9 cis-11 CLA. The cis-9 trans-11 CLA exhibited the highest enrichment
(30.65%), followed by enrichments from 21.06 to 23.08% for trans-10 cis-12, cis-10 trans-12, trans-9
trans-11, and trans-10 trans-12 CLA. The remaining two CLA (cis-9 cis-11 and cis-10 cis-12 CLA) exhibited
enrichments of 18.38 and 19.29%, respectively. The results of this study verified the formation of cis-9
trans-11 and trans-10 cis-12 CLA isomers from linoleic acid biohydrogenation. An additional five CLA isomers
also contained carbons originating from linoleic acid, indicating that pathways of linoleic acid biohydrogenation
are more complex than previously described.
Research Support, Non-U.S. Gov'ts
- Diversity of Cold-Active Protease-Producing Bacteria from Arctic Terrestrial and Marine Environments Revealed by Enrichment Culture
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Eun Hye Kim , Kyeung Hee Cho , Yung Mi Lee , Joung Han Yim , Hong Kum Lee , Jang-Cheon Cho , Soon Gyu Hong
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J. Microbiol. 2010;48(4):426-432. Published online August 20, 2010
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DOI: https://doi.org/10.1007/s12275-010-0015-z
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40
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22
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Abstract
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A new approach for enrichment culture was applied to obtain cold-active protease-producing bacteria for marine and terrestrial samples from Svalbard, Norway. The method was developed for the enrichment of bacteria by long-term incubation at low temperatures in semi-solid agar medium containing meat pieces as the main source of carbon and energy. ZoBell and 0.1× nutrient broth were added for marine and terrestrial microorganisms, respectively, to supply basal elements for growth. One to three types of colonies were observed from each enrichment culture, indicating that specific bacterial species were enriched during the experimental conditions. Among 89 bacterial isolates, protease activity was observed from 48 isolates in the screening media containing skim milk. Good growth was observed at 4°C and 10°C while none of the isolates could grow at 37°C. At low temperatures, enzyme activity was equal to or higher than activity at higher temperatures. Bacterial isolates were included in the genera Pseudoalteromonas (33 isolates), Arthrobacter (24 isolates), Pseudomonas (16 isolates), Psychrobacter (6 isolates), Sphingobacterium (6 isolates), Flavobacterium (2 isolates), Sporosarcina (1 isolate), and Stenotrophomonas (1 isolate). Protease activity was observed from Pseudoalteromonas (33 isolates), Pseudomonas (10 isolates), Arthrobacter (4 isolates), and Flavobacterium (1 isolate).
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- Endophytic Bacterial Diversity in Grapevine (Vitis vinifera L.) Leaves Described by 16S rRNA Gene Sequence Analysis and Length Heterogeneity-PCR
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Daniela Bulgari , Paola Casati , Lorenzo Brusetti , Fabio Quaglino , Milena Brasca , Daniele Daffonchio , Piero Attilio Bianco
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J. Microbiol. 2009;47(4):393-401. Published online September 9, 2009
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DOI: https://doi.org/10.1007/s12275-009-0082-1
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Abstract
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Diversity of bacterial endophytes associated with grapevine leaf tissues was analyzed by cultivation and cultivation-independent methods. In order to identify bacterial endophytes directly from metagenome, a protocol for bacteria enrichment and DNA extraction was optimized. Sequence analysis of 16S rRNA gene libraries underscored five diverse Operational Taxonomic Units (OTUs), showing best sequence matches with gamma-Proteobacteria, family Enterobacteriaceae, with a dominance of the genus Pantoea. Bacteria isolation through cultivation revealed the presence of six OTUs, showing best sequence matches with Actinobacteria, genus Curtobacterium, and with Firmicutes genera Bacillus and Enterococcus. Length Heterogeneity-PCR (LH-PCR) electrophoretic peaks from single bacterial clones were used to setup a database representing the bacterial endophytes identified in association with grapevine tissues. Analysis of healthy and phytoplasma- infected grapevine plants showed that LH-PCR could be a useful complementary tool for examining the diversity of bacterial endophytes especially for diversity survey on a large number of samples.
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