• Title/Summary/Keyword: RNA sequencing analysis

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Microbiological Characteristics of Gouda Cheese Manufactured with Pasteurized and Raw Milk during Ripening Using Next Generation Sequencing

  • Park, Wonseo;Yoo, Jayeon;Oh, Sangnam;Ham, Jun-sang;Jeong, Seok-geun;Kim, Younghoon
    • Food Science of Animal Resources
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    • v.39 no.4
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    • pp.585-600
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    • 2019
  • Gouda cheese, one of most popular cheeses in the Korea, has been produced from only pasteurized milk in Korean dairy farms. Recently, it has become legally possible to produce ripened cheese manufactured with raw milk in Korea. In the present study, we investigated the physico-chemical and microbiological characteristics of Gouda cheese manufactured with raw (R-GC) or pasteurized milk (P-GC) during manufacturing and ripening. Particularly, this study characterized the bacterial community structure of two cheese types, which are produced without pasteurization during ripening based on next generation sequencing of 16S rRNA gene amplicons. During ripening, protein and fat content increased slightly, whereas moisture content decreased in both P-GC and R-GC. At the 6 wk of ripening, R-GC became softer and smoother and hence, the values of hardness and gumminess, chewiness in R-GC was lower than that of P-GC. Metagenomic analysis revealed that the bacterial genera used a starter cultures, namely Lactococcus and Leuconostoc were predominant in both P-GC and R-GC. Moreover, in R-GC, the proportion of coliform bacteria such as Escherichia, Leclercia, Raoultella, and Pseudomonas were detected initially but not during ripening. Taken together, our finding indicates the potential of manufacturing with Gouda cheese from raw milk and the benefits of next generation sequencing for microbial community composition during cheese ripening.

Construction of a Transcriptome-Driven Network at the Early Stage of Infection with Influenza A H1N1 in Human Lung Alveolar Epithelial Cells

  • Chung, Myungguen;Cho, Soo Young;Lee, Young Seek
    • Biomolecules & Therapeutics
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    • v.26 no.3
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    • pp.290-297
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    • 2018
  • We aimed to understand the molecular changes in host cells that accompany infection by the seasonal influenza A H1N1 virus because the initial response rapidly changes owing to the fact that the virus has a robust initial propagation phase. Human epithelial alveolar A549 cells were infected and total RNA was extracted at 30 min, 1 h, 2 h, 4 h, 8 h, 24 h, and 48 h post infection (h.p.i.). The differentially expressed host genes were clustered into two distinct sets of genes as the infection progressed over time. The patterns of expression were significantly different at the early stages of infection. One of the responses showed roles similar to those associated with the enrichment gene sets to known 'gp120 pathway in HIV.' This gene set contains genes known to play roles in preventing the progress of apoptosis, which infected cells undergo as a response to viral infection. The other gene set showed enrichment of 'Drug Metabolism Enzymes (DMEs).' The identification of two distinct gene sets indicates that the virus regulates the cell's mechanisms to create a favorable environment for its stable replication and protection of gene metabolites within 8 h.

Occurrence of Stolbur Phytoplasma Disease in Spreading Type Petunia hybrida Cultivars in Korea

  • Chung, Bong Nam;Jeong, Myeong Il;Choi, Seung Kook;Joa, Jae Ho;Choi, Kyeong San;Choi, In Myeong
    • The Plant Pathology Journal
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    • v.29 no.4
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    • pp.465-470
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    • 2013
  • In January 2012, spreading type petunia cv. Wave Pink plants showing an abnormal growth habit of sprouting unusual multiple plantlets from the lateral buds were collected from a greenhouse in Gwacheon, Gyeonggi Province, Korea. The presence of phytoplasma was investigated using PCR with the primer pairs P1/P6, and R16F1/R1 for nested-PCR. In the nested PCR, 1,096 bp PCR products were obtained, and through sequencing 12 Pet-Stol isolates were identified. Comparison of the nucleotide sequences of 16S rRNA gene of the 12 Pet-Stol isolates with other phytoplasmas belonging to aster yellows or Stolbur showed that Pet-Stol isolates were members of Stolbur. The presence of phytoplasma in petunia was also confirmed by microscopic observation of the pathogens. In this study, Stolbur phytoplasma was identified from spreading type petunia cultivars by sequence analysis of 16S rRNA gene of phytoplasma and microscopic observation of phytoplasma bodies. This is the first report of Stolbur phytoplasma in commercial Petunia hybrida cultivars.

Immune Regulatory Effect of Newly Isolated Lactobacillus delbrueckii from Indian Traditional Yogurt

  • Hong, Yi-Fan;Lee, Yoon-Doo;Park, Jae-Yeon;Jeon, Boram;Jagdish, Deepa;Jang, Soojin;Chung, Dae Kyun;Kim, Hangeun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.8
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    • pp.1321-1323
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    • 2015
  • Lactic acid bacteria (LAB) are microorganisms that are believed to provide health benefits. Here, we isolated LAB from Indian fermented foods, such as traditional Yogurt and Dosa. LAB from Yogurt most significantly induced TNF-α and IL-1β production, whereas LAB from Dosa induced mild cytokine production. After 16S rRNA gene sequencing and phylogenetic analysis, a Yogurt-borne lactic acid bacterium was identified and classified as Lactobacillus delbrueckii subsp. bulgaricus, and it was renamed L. delbrueckii K552 for the further studies. Our data suggest that the newly isolated L. delbrueckii can be used for the treatment of immune deficiency disorders.

Overexpression of FGFR3 mRNA and Mutational Analysis of FGFR3 Gene in Hepatocellular Carcinoma (간암에서 FGFR3 mRNA의 과발현과 FGFR3 유전자의 돌연변이 분석)

  • Chang, Young Gyoon;Bae, Hyun Jin;Nam, Suk Woo
    • YAKHAK HOEJI
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    • v.56 no.6
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    • pp.352-357
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    • 2012
  • FGFR3 is a member of the fibroblast growth factor receptor family which interacts with fibroblast growth factors, setting in motion a cascade of downstream signals, ultimately influencing mitogenesis and differentiation. This particular family member binds acidic and basic fibroblast growth hormone and plays a role in bone development and maintenance. Accumulated evidence suggests that aberrant regulation of FGFR3 and genetic alterations are implicated in the development and progression of various cancers. Despite a high incidence of FGFR3 over-expression, no such investigation has been performed in hepatocellular carcinoma. Thus, we investigated genetic alterations of the FGFR3 gene in 73 cases of hepatocellular carcinoma by single-strand conformational polymorphism (SSCP) and sequencing. One silent mutation (A369A) was found in the extracellular domain of FGFR3, and one genetic alteration in the immunoglobulin-like III domain of FGFR3 appeared to be polymorphism. Taken together, we concluded that over-expression of FGFR3 in hepatocellular carcinoma is not associated with genetic alterations of FGFR3 gene, and we suggest that there could be another underlying mechanism of aberrant FGFR3 expression in hepatocellular carcinoma.

A protein interactions map of multiple organ systems associated with COVID-19 disease

  • Bharne, Dhammapal
    • Genomics & Informatics
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    • v.19 no.2
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    • pp.14.1-14.6
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    • 2021
  • Coronavirus disease 2019 (COVID-19) is an on-going pandemic disease infecting millions of people across the globe. Recent reports of reduction in antibody levels and the re-emergence of the disease in recovered patients necessitated the understanding of the pandemic at the core level. The cases of multiple organ failures emphasized the consideration of different organ systems while managing the disease. The present study employed RNA sequencing data to determine the disease associated differentially regulated genes and their related protein interactions in several organ systems. It signified the importance of early diagnosis and treatment of the disease. A map of protein interactions of multiple organ systems was built and uncovered CAV1 and CTNNB1 as the top degree nodes. A core interactions sub-network was analyzed to identify different modules of functional significance. AR, CTNNB1, CAV1, and PIK3R1 proteins were unfolded as bridging nodes interconnecting different modules for the information flow across several pathways. The present study also highlighted some of the druggable targets to analyze in drug re-purposing strategies against the COVID-19 pandemic. Therefore, the protein interactions map and the modular interactions of the differentially regulated genes in the multiple organ systems would incline the scientists and researchers to investigate in novel therapeutics for the COVID-19 pandemic expeditiously.

Identification of Vibrio species isolated from cultured olive flounder (Paralichthys olivaceus) in Jeju Island, South Korea

  • Sohn, Hanchang;Kim, Jeongeun;Jin, Changnam;Lee, Jehee
    • Fisheries and Aquatic Sciences
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    • v.22 no.7
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    • pp.14.1-14.8
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    • 2019
  • Olive flounder (Paralichthys olivaceus) is the major species developed for aquaculture in South Korea. Over the long history of olive flounder aquaculture, complex and diverse diseases have been a major problem, negatively impacting industrial production. Vibriosis is a prolific disease which continuously damages olive flounder aquaculture. A bacterial disease survey was performed from January to June 2017 on 20 olive flounder farms on Jeju Island. A total of 1710 fish were sampled, and bacteria from the external and internal organs of 560 fish were collected. Bacterial strains were identified using 16 s rRNA sequencing. Twenty-seven species and 184 strains of Vibrio were isolated during this survey, and phylogenetic analysis was performed. Bacterial isolates were investigated for the distribution of pathogenic and non-pathogenic species, as well as bacterial presence in tested organs was characterized. V. gigantis and V. scophthalmi were the dominant non-pathogenic and pathogenic strains isolated during this survey, respectively. This study provides data on specific Vibrio spp. isolated from cultured olive flounder in an effort to provide direction for future research and inform aquaculture management practices.

Discrepancies between Mitochondrial DNA and AFLP Genetic Variation among Lineages of Sea Slaters Ligia in the East Asian Region

  • Kang, Seunghyun;Jung, Jongwoo
    • Animal Systematics, Evolution and Diversity
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    • v.36 no.4
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    • pp.347-353
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    • 2020
  • Although sea slaters Ligia have a significant role in rocky shore habitats, their taxonomic entities have not been clearly understood. In this study, we investigated whether genetic variation inferred from a nuclear genetic marker, namely amplified fragment length polymorphism (AFLP), would conform to that of a mitochondrial DNA marker. Using both the mitochondrial DNA marker and the AFLP marker amplified by the six selective primer sets, we analyzed 95 Ligia individuals from eight locations from East Asia. The direct sequencing of mitochondrial 16S rRNA gene revealed three distinct genetic lineages, with 9.8-11.7 Kimura 2-parameter genetic distance. However, the results of AFLP genotyping analysis with 691 loci did not support those of mitochondrial DNA, and revealed an unexpectedly high proportion of shared polymorphisms among lineages. The inconsistency between the two different genetic markers may be explained by difference in DNA evolutionary history, for example inheritance patterns, effective population size, and mutation rate. The other factor is a possible genomic island of speciation, in that most of the genomic parts are shared among lineages, and only a few genomic regions have diverged.

Overlapping Region of p53/Wrap53 Transcripts: Mutational Analysis and Sequence Similarity with microRNA-4732-5p

  • Pouladi, Nasser;Kouhsari, Shideh Montasser;Feizi, Mohammadali Hosseinpour;Gavgani, Reyhaneh Ravanbakhsh;Azarfam, Parvin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3503-3507
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    • 2013
  • Background: Although the majority of investigations concerned with TP53 and its protein have focused on coding regions, recently a set of studies highlighted significant roles of regulatory elements located in p53 mRNA, especially 5'UTR. The wrap53${\alpha}$ transcript is one of those that acts as a natural antisense agent, forming RNA-RNA hybrids with p53 mRNA and protecting it from degradation. Materials and Methods: In this study, we focused on the mutation status of exon $1{\alpha}$ of the WRAP53 gene (according to exon 1 of p53) in 160 breast tumor tissue samples and conducted a bioinformatics search for probable miRNA binding site in the p53/wrap53 overlapping region. Mutations were detected, using single stranded conformation polymorphism (SSCP) and sequencing. We applied the miRBase database for prediction of miRNAs which target overlapping region of p53/wrap53 transcripts. Results: Our results showed all samples to have wild type alleles in exon 1 of TP53 gene. We could detect a novel and unreported intronic mutation (IVS1+56, G>C) outside overlapping regions of p53/wrap53 genes in breast cancer tissues and also predict the presence of a binding site for miR-4732-5p in the 5'UTR of Wrap53 mRNA. Conclusions: From our findings we propose designing further studies focused on overexpression of miRNA-4732-5p and introducing different mutations in the overlapping region of wrap53 and p53 genes in order to study their effects on p53 and its ${\Delta}N$ isoform (${\Delta}$40p53) expression. The results may provide new pieces in the p53 targeting puzzle for cancer therapy.

Analysis of Microbial Community Change in Ganjang According to the Size of Meju (메주의 크기에 따른 간장의 미생물 군집 변화 양상 분석)

  • Ho Jin Jeong;Gwangsu Ha;Ranhee Lee;Do-Youn Jeong;Hee-Jong Yang
    • Journal of Life Science
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    • v.34 no.7
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    • pp.453-464
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    • 2024
  • The fermentation of ganjang is known to be greatly influenced by the microbial communities derived from its primary ingredients, meju and sea salt. This study investigated the effects of changes in meju size on the distribution and correlation of microbial communities in ganjang fermentation, to enhance its fermentation process. Ganjang was prepared using whole meju and meju divided into thirds, and samples were collected at 7-day intervals over a period of 28 days for microbial community analysis based on 16S rRNA gene sequencing. At the genus level, during fermentation, ganjang made with whole meju exhibited a dominance of Chromohalobacter (day 7), Pediococcus (day 14), Bacillus (day 21), and Pediococcus (day 28), whereas ganjang made with meju divided into thirds consistently showed a Pediococcus predominance over the 28 days. Beta-diversity analysis of microbial communities in ganjang with different meju sizes revealed significant separation of microbial communities at fermentation days 7 and 14 but not at days 21 and 28 across all experimental groups. The linear discriminant analysis effect size (LEfSe) was determined to identify biomarkers contributing to microbial community differences at days 7 and 14, showing that on day 7, potentially halophilic microbes such as Gammaproteobacteria, Firmicutes, Oceanospirillales, Halomonadaceae, Bacilli, and Chromohalobacter were prominent, whereas on day 14, lactic acid bacteria such as Pediococcus acidilactici, Lactobacillaceae, Pediococcus, Bacilli, Leuconostocaceae, and Weissella were predominant. Furthermore, correlation analysis of microbial communities at the genus and species levels revealed differences in correlation patterns between meju sizes, suggesting that meju size may influence microbial interactions within ganjang.