• Title/Summary/Keyword: 차세대염기서열 분석

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Comparative Microbiome Analysis of and Microbial Biomarker Discovery in Two Different Fermented Soy Products, Doenjang and Ganjang, Using Next-generation Sequencing (차세대 염기서열 분석법을 이용한 된장과 간장의 미생물 분포 및 바이오마커 분석)

  • Ha, Gwangsu;Jeong, Ho Jin;Noh, Yunjeong;Kim, JinWon;Jeong, Su-Ji;Jeong, Do-Youn;Yan, Hee-Jong
    • Journal of Life Science
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    • v.32 no.10
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    • pp.803-811
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    • 2022
  • Despite the importance of traditional Korean fermented foods, little is known about the microbial communities and diversity of fermented soy products. To gain insight into the unexplored microbial communities of both Doenjang (DJ) and Ganjang (GJ) that may contribute to the fermentation in Korean traditional foods, we carried out next-generation sequencing (NGS) based on the V3-V4 region of 16S rDNA gene analysis. The alpha diversity analysis results revealed that both the Shannon and Simpson diversity indices were significantly different between the two groups, whereas the richness indices, including ACE, CHAO, and Jackknife, were not significant. Firmicutes were the most dominant phylum in both groups, but several taxa were found to be more abundant in DJ than in GJ. The proportions of Bacillus, Kroppenstedtia, Clostridium, and Pseudomonas and most halophiles and halotolerant bacteria, such as Tetragenococcus, Chromohalobacter, Lentibacillus, and Psychrobacter, were lower in DJ than in GJ. Linear discriminant effect size (LEfSe) analysis was carried out to discover discriminative functional biomarkers. Biomarker discovery results showed that Bacillus and Tetragenococcus were identified as the most important features for the classification of subjects to DJ and GJ. Paired-permutational multivariate analysis of variance (PERMANOVA) further revealed that the bacterial community structure between the two groups was statistically different (p=0.001).

Imputation Accuracy from 770K SNP Chips to Next Generation Sequencing Data in a Hanwoo (Korean Native Cattle) Population using Minimac3 and Beagle (Minimac3와 Beagle 프로그램을 이용한 한우 770K chip 데이터에서 차세대 염기서열분석 데이터로의 결측치 대치의 정확도 분석)

  • An, Na-Rae;Son, Ju-Hwan;Park, Jong-Eun;Chai, Han-Ha;Jang, Gul-Won;Lim, Dajeong
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1255-1261
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    • 2018
  • Whole genome analysis have been made possible with the development of DNA sequencing technologies and discovery of many single nucleotide polymorphisms (SNPs). Large number of SNP can be analyzed with SNP chips, since SNPs of human as well as livestock genomes are available. Among the various missing nucleotide imputation programs, Minimac3 software is suggested to be highly accurate, with a simplified workflow and relatively fast. In the present study, we used Minimac3 program to perform genomic missing value substitution 1,226 animals 770K SNP chip and imputing missing SNPs with next generation sequencing data from 311 animals. The accuracy on each chromosome was about 94~96%, and individual sample accuracy was about 92~98%. After imputation of the genotypes, SNPs with R Square ($R^2$) values for three conditions were 0.4, 0.6, and 0.8 and the percentage of SNPs were 91%, 84%, and 70% respectively. The differences in the Minor Allele Frequency gave $R^2$ values corresponding to seven intervals (0, 0.025), (0.025, 0.05), (0.05, 0.1), (0.1, 0.2), (0.2, 0.3). (0.3, 0.4) and (0.4, 0.5) of 64~88%. The total analysis time was about 12 hr. In future SNP chip studies, as the size and complexity of the genomic datasets increase, we expect that genomic imputation using Minimac3 can improve the reliability of chip data for Hanwoo discrimination.

Analyzing Vomit of Platalea minor (Black-faced Spoonbill) to Identify Food Components using Next-Generation Sequencing and Microscopy (차세대염기서열 및 현미경 분석을 통한 저어새의 토사물 내 먹이생물 분석)

  • Kim, Hyun-Jung;Lee, Taek-Kyun;Jung, Seung Won;Kwon, In-Ki;Yoo, Jae-Won
    • Korean Journal of Environmental Biology
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    • v.36 no.2
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    • pp.165-173
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    • 2018
  • We sampled vomit of black-faced spoonbills(Platalea minor) during the brood-rearing season (from June 2011 to June 2014) at the Namdong reservoir in Incheon and analyzed the food components in the vomit using microscopy and next-generation sequencing (NGS). Microscopic observations primarily helped in identifying osteichthyes (bony fishes), crustaceans, and polychaetes. In particular, species belonging to the families Mugilidae and Gobiidae among the fish, and Macrophthalmus japonicas among the crustaceans, were observed at high frequency. Results of NGS analysis revealed the predominant presence of bony fish (42.58% of total reads) and crustaceans (40.75% of total reads), whereas others, such as polychaetes (12.66%), insects (0.24%), and unidentified species (3.78%), occurred in lower proportions. At the species level, results of NGS analysis revealed that Macrophthalmus abbreviates and Macrobrachium sp. among the crustaceans, and Acanthogobius hasta, Tridentiger obscurus, and Pterogobius zacalles among the bony fish, made up a high proportion of the total reads. These food species are frequently found at tidal flats in the Songdo and Sihwa lakes, emphasizing the importance of these areas as potential feeding sites of the black-faced spoonbill. Feed composition of the black-faced spoonbill, as evaluated by analyzing its vomit, differed when the evaluations were done by microscopic observation or by NGS analysis. Evaluation by microscopic observation is difficult and not error free, owing to the degradation of the samples to be analyzed; however, NGS analysis is more accurate, because it makes use of genetic information. Therefore, accurately analyzing food components from morphologically indistinguishable samples is possible by using genetic analysis.

Genomics and Molecular Markers for Major Cucurbitaceae Crops (주요 박과작물의 유전체 및 분자마커 연구 현황)

  • Park, Girim;Kim, Nahui;Park, Younghoon
    • Journal of Life Science
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    • v.25 no.9
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    • pp.1059-1071
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    • 2015
  • Watermelon and melon are economically important Cucurbitaceae crops. Recently, the development of molecular markers based on the construction of genetic linkage maps and detection of DNA sequence variants through next generation sequencing are essential as molecular breeding strategies for crop improvement that uses marker-assisted selection and backcrossing. In this paper, we intended to provide useful information for molecular breeding of watermelon and melon by analyzing the current status of international and domestic research efforts on genomics and molecular markers. Due to diverse genetic maps constructed and the reference genome sequencing completed in the past, DNA markers that are useful for selecting important traits including yield, fruit quality, and disease resistances have been reported and publicly available. To date, more than 16 genetic maps and loci and linked markers for more than 40 traits have reported for each watermelon and melon. Furthermore, the functional genes that are responsible for those traits are being continuously discovered by high-density genetic map and map-based cloning. In addition, whole genome resequencing of various germplasm is under progress based on the reference genome. Not only by the efforts for developing novel molecular markers, but application of public marker information currently available will greatly facilitate breeding process through genomics-assisted breeding.

An Efficient Parallelization Mechanism for Preprocessing of Genome Sequence Data on HPC environment (고성능 클러스터와 분산 병렬 파일 시스템을 이용한 유전체데이터 전처리 작업의 효율적인 병렬화 기법)

  • Byun, Eun-Kyu;Mun, Ji-hyeob;Kwak, Jae-Hyuck
    • Proceedings of the Korea Information Processing Society Conference
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    • 2018.10a
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    • pp.50-53
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    • 2018
  • 차세대 염기서열 분석법이 생성한 유전체 원시 데이터를 기존의 방식대로 하나의 서버에서 분석하기 위해서는 수십 시간이 필요할 수 있고 이러한 시간을 최대한 줄여야 하는 응급 상황도 존재한다. 따라서 본 연구에서는 고속의 네트워크로 연결되고 병렬 파일 시스템을 공유하는 서버 클러스터를 활용하여 분석 시간을 크게 단축 시킬 수 있는 유전체 데이터 분석의 전처리 프로세스의 병렬화 방법을 제안한다. 기존의 검증된 분석도구를 기반으로 프로세스의 병렬화, 데이터의 분배 및 병렬 병합 기법을 개발하였고 실험을 통해 성능을 향상 시킬 수 있음을 증명하였다.

A Study on the Hierarchical Expression of Human Cell Lineage (인간 세포 Lineage 의 계층적 표현에 관한 연구)

  • Park, JaeSoon;Kwon, Seong Gyu;Oh, Ji Won;Lee, JongHyuk
    • Proceedings of the Korea Information Processing Society Conference
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    • 2020.11a
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    • pp.663-664
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    • 2020
  • 차세대 염기서열 분석 기술은 성능과 비용 면에서 매우 향상되어 한 개체 내 여러 세포의 유전자 분석이 가능한 수준이다. 한 개체 내 여러 조직 세포의 유전자는 모두 동일하지 않기 때문에 여러 조직 세포의 Lineage 를 계층적으로 표현하고 이를 조직 세포 간 변이 정도를 파악하는 데 활용한다면 암 돌연변이 발생 등을 미리 예측할 수 있다. 본 논문은 한 개체 내 여러 조직 간 변이를 관찰하기 위해 변이 검출 데이터를 계층적 군집 방법을 이용해 분석하고 이를 시각화 하는 방법을 제안한다. 실제의 8 개 조직 세포의 유전자를 분석하고 변이를 검출하여 Dendrogram 그래프로 시각화 하였다.

A Study on the Induction of Infertility of Largemouth Bass (Micropterus salmoides) by CRISPR/Cas9 System (CRISPR/Cas9 System을 활용한 배스의 불임 유도에 대한 연구)

  • Park, Seung-Chul;Kim, Jong Hyun;Lee, Yoon Jeong
    • Korean Journal of Environment and Ecology
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    • v.35 no.5
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    • pp.503-524
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    • 2021
  • A largemouth bass (Micropterus salmoides) is an ecosystem disturbance fish species at the highest rank in the aquatic ecosystem, causing a serious imbalance in freshwater ecosystems. Although various attempts have been made to eradicate and control largemouth bass, no effective measures were found. Therefore, it is necessary to find an approach to maximize the effective population reduction based on the unique characteristics of largemouth bass. This study used the transcriptome analysis to derive 182,887 unigene contigs and select 12 types of final target sequences for applying the CRISPR/Cas9 system in the genes of IZUMO1 and Zona pellucida sperm-binding protein, which are proteins involved in sperm-egg recognition. After synthesizing 12 types of sgRNA capable of recognizing each target sequence, 12 types of Cas9-sgRNA ribonucleoprotein (RNP) complexes to be used in subsequent studies were prepared. This study searched the protein-coding gene of sperm-egg through the Next Generation Sequencing (NGS) and edited genes through the CRISPR/Cas9 system to induce infertile individuals that produced reproductive cells but could not form fertilized eggs. Through such a series of processes, it successfully established a composition development process for largemouth bass. It is judged that this study contributed to securing the valuable basic data for follow-up studies to verify its effect for the management of ecological disturbances without affecting the habitat of other endemic species in the same water system with the largemouth bass.

Comparison of Microbial Community Compositions between Doenjang and Cheonggukjang Using Next Generation Sequencing (차세대 염기서열 분석법을 이용한 전통 된장과 청국장의 미생물 분포 분석)

  • Ha, Gwangsu;Kim, JinWon;Shin, Su-Jin;Jeong, Su-Ji;Yang, Hee-Jong;Jeong, Do-Youn
    • Journal of Life Science
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    • v.31 no.10
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    • pp.922-928
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    • 2021
  • To profile the microbial compositions of Korean traditional fermented paste made from whole soybeans, Doenjang and Cheonggukjang, and compare their taxonomic differences, we analyzed the V3-V4 region of 16S rRNA of naturally fermented foods by using next generation sequencing. α-Diversity results showed that values indicating bacterial community abundances (OTUs) and richness (ACE, Chao1) were statistically significant (p=0.0001) in Doenjang and Cheonggukjang. Firmicutes was the most common phylum in both groups, representing 97.02% and 99.67% in the Doenjang and Cheonggukjang groups, respectively. Bacillus was the most dominant genus, accounting for 71.70% and 59.87% in both groups. Linear discriminant (LDA) effect size (LEfSe) analysis was performed to reveal the significant ranking of abundant taxa in different fermented foods. A size-effect threshold of 2.0 on the logarithmic LDA score was used for discriminative functional biomarkers. On the species level, Bacillus subtilis, Tetragenococcus halophilus, and Clostridium arbusti were significantly more abundant in Doenjang than in Cheonggukjang, whereas Bacillus thermoamylovorans, Enterococcus faecium, and Lactobacillus sakei were significantly more abundant in Cheonggukjang than in Doenjang. Permutational multivariate analysis of variance (PERMANOVA) showed that the statistical difference in microbial clusters between the two groups was significant at the confidence level of p=0.001. This research could be used as basic research to identify the correlation between the biochemical characteristics of Korean fermented foods and the distribution of microbial communities.

Development of SNP markers for the identification of apple flesh color based on RNA-Seq data (RNA-Seq data를 이용한 사과 과육색 판별 SNP 분자표지 개발)

  • Kim, Se Hee;Park, Seo Jun;Cho, Kang Hee;Lee, Han Chan;Lee, Jung Woo;Choi, In Myung
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.372-378
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    • 2017
  • For comparison of the transcription profiles in apple (Malus domestica L.) cultivars differing in flesh color expression, two cDNA libraries were constructed. Differences in gene expression between red flesh apple cultivar, 'Redfield' and white flesh apple cultivar, 'Granny Smith' were investigated by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the red flesh apple cultivar and white flesh apple cultivar were selected for nucleotide sequence determination and homology searches. High resolution melting (HRM) technique measures temperature induced strand separation of short PCR amplicons, and is able to detect variation as small as one base difference between red flesh apple cultivars and white flesh apple cultivars. We applied high resolution melting (HRM) analysis to discover single nucleotide polymorphisms (SNP) based on the predicted SNP information derived from the apple EST database. All 103 pairs of SNPs were discriminated, and the HRM profiles of amplicons were established. Putative SNPs were screened from the apple EST contigs by HRM analysis displayed specific difference between 10 red flesh apple cultivars and 11 white flesh apple cultivars. In this study, we report an efficient method to develop SNP markers from an EST database with HRM analysis in apple. These SNP markers could be useful for apple marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in apple cultivars.