• Korean Journal of Clinical Laboratory Science
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• v.55 no.4
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• pp.235-243
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• 2023

As next-generation sequencing has been developed and used widely, RNA-sequencing (RNA-seq) has rapidly emerged as the first choice of tools to validate global transcriptome profiling. With the significant advances in RNA-seq, various types of RNA-seq have evolved in conjunction with the progress in bioinformatic tools. On the other hand, it is difficult to interpret the complex data underlying the biological meaning without a general understanding of the types of RNA-seq and bioinformatic approaches. In this regard, this paper discusses the two main sections of RNA-seq. First, two major variants of RNA-seq are described and compared with the standard RNA-seq. This provides insights into which RNA-seq method is most appropriate for their research. Second, the most widely used RNA-seq data analyses are discussed: (1) exploratory data analysis and (2) pathway enrichment analysis. This paper introduces the most widely used exploratory data analysis for RNA-seq, such as principal component analysis, heatmap, and volcano plot, which can provide the overall trends in the dataset. The pathway enrichment analysis section introduces three generations of pathway enrichment analysis and how they generate enriched pathways with the RNA-seq dataset.

• Research in Plant Disease
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• v.30 no.1
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• pp.60-65
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• 2024

In July 2020, total RNA was extracted from passion fruit (Passiflora edulis) leaves showing virus symptoms such as chlorotic spots and vein banding in Haenam, South Korea. Cucumber mosaic virus (CMV)-HN2 was identified through reverse transcription polymerase chain reaction and sequencing analysis. To confirm the biological characteristics of the CMV infecting passion fruit, 10 indicator plants were inoculated with CMV-HN2, and the results showed a typical CMV symptoms. Phylogenetic analysis based on the amino acid of the coat protein (CP) of CMVs revealed that the CMV passion fruit isolates belonged to subgroup I, among which CMV-HN2 belonged to subgroup IA. Additionally, CMVs isolated from passion fruit in Korea have amino acid sequence variation between the subgroup. Among them, CMV-HN2 had four to eight amino acid differences in CP from other CMV isolates from passion fruit. These results confirm the presence of genetic diversity in the CPs of passion fruit CMV isolates.

• Journal of Life Science
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• v.34 no.10
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• pp.691-700
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• 2024

Frutobacillus spp. strains (Fs) were isolated from honeybee larvae to evaluate their functionality and potential use as probiotics. Anti-microbial activity was generally observed in F strains against Gram-negative, Gram-positive, and yeast strains, with F2 and F3 being superior, particularly F2. Kerosene emulsification was similarly observed in strains other, except for F4. Emulsification analysis based on carbon sources showed that F2 had high emulsification in the presence of fructose but lower than the standard strain and 16S rDNA sequence analysis revealed that the F2 was identified as Fructobacillus fructosus. The growth curve of F2 showed maximum growth at 18 hr, followed by a slight increase. Furthermore, antimicrobial activity and pH showed maximum and minimum values at 18 hr, respectively, and remained constant thereafter. Lactic acid content showed a slight decrease after reaching its maximum value at 24 hr of culture. Acid resistance was observed up to pH 2.5, but completely lost at pH 2.0. Bile acid resistance was generally strong. F2's adhesion to mucin was higher compared to S. Typhimurium, which increased until 18 hr of culture and then decreased. Enzyme activity according to anaerobic culture time was high for carboxymethylcellulase (CMCase), avicelase, and mannase, regardless of the presence of fructose. Auto- and co-aggregation were higher compared to the standard strain, and surface hydrophobicity was high for chloroform, which indicates electron donor properties. Therefore, the Fructobacillus fructosus F2 can be considered a potential probiotic due to its excellent anti-microbial, emulsification, acid resistance, bile resistance, CMCase, mannase, and auto- and co-aggregation properties.

• Journal of Soil and Groundwater Environment
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• v.10 no.2
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• pp.52-58
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• 2005

In Korea, little attention has been paid to microbial perchloroethylene (PCE) and/or trichloroethylene (TCE) dechlorination activity and identification of microorganisms involved in PCE reductive dechlorination at a PCE-contaminated aquifer. We performed microcosm tests using the groundwater samples from 4 different contaminated sites (i.e. Changwon A, Changwon B, Bucheon and Yangsan) to assess PCE reductive dechlorination activity. We also adapted molecular techniques to screen what types of known reductive dechlorinators are present at the PCE-contaminated aquifers. In the Changwon A and Changwon B active microcosms where potential electron donors such as sodium propionate, sodium lactate, sodium butyrate, and sodium fumarate, were added, ethylene, an end-product of complete reductive dechlorination of PCE, was detected after a period of 90 days of incubation. In the Bucheon and Yangsan active microcosms, cis-1,2-dichloroethylene (c-DCE) was accumulated without the production of vinyl chloride (VC) and ethylene. Molecular techniques were used to evaluate the microbial community structures in the Changwon B and Yangsan aquifer. We found two sequence types that were closely related to a known PCE to ethylene dechlorinator, named uncultured bacterium clone DCE47, in the Changwon B site clone library. However, in the Yangsan site clone library, no sequence type was closely related to known PCE dechlorinators reported. It is plausible that microorganisms being capable of completely dechlorinating PCE to ethylene may be present in the Changwon B site aquifer. In this study we find that complete PCE reductive dechlorinators are present at some PCE-contaminated sites in Korea. In an engineering point of view this information makes it feasible to apply a biological reductive dechlorination process for remediating PCE- and/or TCE-contaminated aquifers in Korea.

• The Korean Journal of Malacology
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• v.32 no.4
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• pp.263-268
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• 2016

Metallothionein (MT) family of metal-binding proteins are involved in maintaining homeostasis and heavy metal poisoning. Recently, MT has been considered as a biomarker that can identify a particular species, very similar to the use of cytochrome oxidase I (COI) gene. Satsuma myomphala species of land snails have been reported from North-East Asia, including South Korea and Japan. In particular, the land snail species have been known from only a limited area of Geoje Island, Gyeongsangnam-do province of South Korea. Genetic studies of S. myomphala has been limited with only 6 nucleotide, 2 protein registered on the NCBI server. For elucidating the genetic information of S. myomphala, we conducted RNA sequencing analysis using Illumina HiSeq 2500 next-generation platform. We screened the MT gene from the RNA-Seq database to confirm the molecular phylogenetic relationship. After sequencing, the de novo analysis and clustering generated 103,774 unigenes. After annotation against PANM database using BLAST program, we obtained MT sequence of 74 amino acid residues containing the coding region of 222 bp. Based on this sequence, we found about 53 sequences using the BLAST program in NCBI nr database. Using ClustalX alignment, Maximum-Likehood Tree of MEGA program, we confirmed the molecular phylogenetic relationships that showed similarity with mollusks such as Helix pomatia and H. aspersa, Megathura crenulata.

• Korean Journal of Microbiology
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• v.49 no.2
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• pp.137-145
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• 2013

This study was performed at 2 sites of Nak-Dong River to investigate the changes of nitrifiers depending on the presence and absence of organic pollutants (due to the effluents of domestic wastewater treatment plant, WWTP). Conventional chemical parameters such as T-N, $NH_4$-N, $NO_2$-N, $NO_3$-N were measured and the quantitative nitrifiers at the 2 sites were analyzed comparatively by fluorescent in situ hybridization (FISH) with NSO190 and NIT3, after checking the presence of gene amoA of ammonia oxidizing bacteria (AOB) and 16S rDNA signature sequence for Nitrobacter sp. that belongs to nitrite oxidizing bacteria (NOB). Also ${\alpha}{\cdot}{\beta}{\cdot}{\gamma}$-Proteobacteria were detected using FISH to get a glimpse of the general bacterial community structure of the sites. Based on the distribution structure of the ${\alpha}{\cdot}{\beta}{\cdot}{\gamma}$-Proteobacteria and the measurement of nitrogen in different phases, it could be said that the site 2 was more polluted with organics than site 1. Corresponding to the above conclusion, the average numbers of AOB and NOB detected by NSO160 and NIT3, respectively, at site 2 [AOB, $9.3{\times}10^5$; NOB, $1.6{\times}10^6$ (cells/ml)] was more than those at site 1 [AOB, $7.8{\times}10^5$; NOB, $0.8{\times}10^6$ (cells/ml)] and also their ratios to total counts were higher at site 2 (AOB, 27%; NOB, 34%) than those at site 1 (AOB, 18%; NOB, 23%). Thus, it could be concluded that the nitrification at site 2 was more active due to continuous loading of organics from the effluents of domestic WWTP, compared to site 1 located closed to raw drinking water supply and subsequently less polluted with organics.

• Korean Journal of Environment and Ecology
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• v.31 no.4
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• pp.349-364
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• 2017

Duckweed family (Lemnaceae Martinov), including the genus Lemna L., is a typical floating aquatic perennial plant, and about five genera and 40 species in the family are in wide distribution around the world except the polar regions. The genus Lemna is the smallest and the simplest plant among the angiosperms. It has a characteristic of doubling every three days with fast vegetative propagation, which helps the organisms to increase in rapid growth. As such, the plant is ideal for environmental pollution assessment and toxicity test. Although taxonomists and scholars have used different scientific names for the species, many of them have agreed that there is only one member of species of the genus Lemna in Korea. Paying attention to the external morphological variation observed in the Korean genus Lemna, we conducted a molecular phylogenetic analysis to identify the entity of the Korean Lemna species and to investigate the possibility of two or more members of the species existing in Korea. We determined and aligned the DNA sequences of the atpF-H region of the chloroplast DNA in 37 populations of the nationally distributed Lemna species. The results showed that the sequence length of the cp DNA atpF-H region was 463-483 bp, the length of the aligned sequences was 488 bp, and the number of variation site in nucleotide sequences was 47. There were two types of aligned sequences of the cp DNA atpF-H region from 37 populations of Lemna species in Korea. The maximum parsimony analysis revealed that the Korean Lemna consists of two clades, and one of them had two subclades. The results suggest that, contrary to the general understanding, at least two taxa (L.aequinoctialis, L.minor) exist in Korea.

• Journal of Applied Biological Chemistry
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• v.52 no.1
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• pp.1-7
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• 2009

For the development of qualitative PCR detection method of genetically modified (CM) rice, rice species-specific gene, OsCc-1 (rice cytochrome c gene), was selected as suitable far use as an endogenous gene in rice. The primer pair OsCytC-5'/3'with 111 bp amplicon was used for PCR amplification of the rice endogenous gene, OsCc-1 and no amplified product was observed from 8 different crops as templates. Qualitative PCR method was carried out with stack traits of L528$\times$CryIAc1 GM rice developed in Korea. For the qualitative PCRs, some primer pairs were designed with a construct-specific and event-specific type based on T-DNA and junction sequences of T-DNA in GM rice. Actck-5'/3' amplifying between actin promoter and OsCK1 gene introduced in LS28 gave rise to an amplicon 306 bp; also, CrLB-5'/3' from CryIAcl and CKRB-5'/3'amplifying the junction region of T-DNA and genome sequence from LS28 as event-specific primers gave rise to an amplicon 142 bp and 91 bp, respectively. These primer pairs for the detection of event-specific targets not produced PCR amplicons on non-CM rice and various crops in contrast to event lines. Therefore, in this study we verified that event-specific primers were effective to specifically detect stack trait lines and demonstrated that this method presented could be provided with the detection-method data for risk assessment analysis of GM rice to be commercialized.

• Applied Biological Chemistry
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• v.50 no.3
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• pp.167-177
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• 2007

This study was carried out to develop natural vinegared kochujang. We first formulated vinegared kochujang containing pine needle (PN) and Artemisia princeps (AP) extracts. A dominant strain was isolated from the vinegared kochujang and identified as Bacillus amyloliquefaciens ER282 based on the 16S rDNA sequence. The plant extracts of PN and AP were also investigated for their antibacterial activity and showed enough good activities to use for vinegared kochujang as a natural preservative. Quality characteristics of vinegared kochujang were evaluated, based on total cell numbers, pH, acidity, sugar content, color and sensory evaluation during storage at $25^{\circ}C$ and $37^{\circ}C$, respectively, for 3 weeks. No significant differences were found in the pHs and acidities of vinegared kochujang added with the plant extracts of PN and AP during their storage compared to starting time; however, total cell numbers were gradually decreased during the storage at $25^{\circ}C$ and $37^{\circ}C$, as the storage period was increased. For sensory evaluation, overall preference continuously declined depending on storage period, but the addition of 3% extracts of PN could delay the quality loss of vinegared kochujang during storage at $25^{\circ}C$ and $37^{\circ}C$ for 3 weeks.

• Applied Biological Chemistry
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• v.50 no.4
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• pp.268-275
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• 2007

Diversity of the mud flat microbial population in Suncheon Bay was investigated by studying extracellular enzyme activities and 16S rDNA sequences. Four culturable bacterial strains with CMCase, xylanase and protease activities were isolated from the wetland and the mud flat. All the strains produced more xylanase activity than CMCase or protease activity, and the properties of the isolate enzymes from the wetland were similar to those from the mud flat. About 2,000 clones were obtained with the 16S rDNA amplified from the metagenomic DNA isolated from the mud samples. Based on the restriction pattern(s), seventeen clones were selected for base sequence analysis. Of the 17 clones, only 35% (6 clones) were found to be cultured strains and 65% (11 clones) to be uncultured strains. The similarities in the base sequences of the clones ranged from 91.0% to 99.9% with an average similarity of 97.3%. The clones could be divided into 7 groups, Proteobacteria (9 clones, 52.9%), Firmicutes (3 clones, 17.6%), Bacteroidetes (1 clone), Flavobacteria (1 clone), Verrucomicrobia (1 clone), Acidobacteria (1 clone), and Chloroflexi (1 clone). Most of the Proteobacteria clones were gamma Proteobacteria associated with oxidation-reduction of sulfur.