• Title/Summary/Keyword: Next-Generation Sequencing

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Development of Solanum hougasii-specific markers using the complete chloroplast genome sequences of Solanum species (엽록체 전장유전체 정보를 이용한 Solanum hougasii 특이적 분자마커 개발)

  • Kim, Soojung;Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.141-149
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    • 2020
  • Solanum hougasii, one of the wild Solanum species, has been widely used in potato breeding since it exhibits excellent resistance to diverse important pathogens. S. hougasii can be directly crossed with the cultivated tetraploid potato (S. tuberosum) owing to its EBN (Endosperm Balanced Number) value of 4, which is same as that of S. tuberosum although it is an allohexaploid. In this study, the complete chloroplast genome sequence of S. hougasii was obtained by next-generation sequencing technology, and compared with that of the chloroplast genome of seven other Solanum species to identify S. hougasii-specific PCR markers. The length of the complete chloroplast genome of S. hougasii was 155,549 bp. The structural organization of the chloroplast genome in S. hougasii was found to be similar to that of seven other Solanum species studied. Phylogenetic analysis of S. hougasii with ten other Solanaceae family members revealed that S. hougasii was most closely related to S. stoloniferum, followed by S. berthaultii, and S. tuberosum. Additional comparison of the chloroplast genome sequence with that of five other Solanum species revealed five InDels and 43 SNPs specific to S. hougasii. Based on these SNPs, four PCR-based markers were developed for the differentiation of S. hougasii from other Solanum species. The results obtained in this study will aid in exploring the evolutionary and breeding aspects of Solanum species.

Transcription Analysis of Recombinant Trichoderma reesei HJ-48 to Compare the Molecular Basis for Fermentation of Glucose and Xylose

  • Huang, Jun;Lin, Mei;Liang, Shijie;Qin, Qiurong;Liao, Siming;Lu, Bo;Wang, Qingyan
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1467-1479
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    • 2020
  • Profiling the transcriptome changes involved in xylose metabolism by the fungus Trichoderma reesei allows for the identification of potential targets for ethanol production processing. In the present study, the transcriptome of T. reesei HJ-48 grown on xylose versus glucose was analyzed using next-generation sequencing technology. During xylose fermentation, numerous genes related to central metabolic pathways, including xylose reductase (XR) and xylitol dehydrogenase (XDH), were expressed at higher levels in T. reesei HJ-48. Notably, growth on xylose did not fully repress the genes encoding enzymes of the tricarboxylic acid and respiratory pathways. In addition, increased expression of several sugar transporters was observed during xylose fermentation. This study provides a valuable dataset for further investigation of xylose fermentation and provides a deeper insight into the various genes involved in this process.

Chromothripsis in Treatment Resistance in Multiple Myeloma

  • Lee, Kyoung Joo;Lee, Ki Hong;Yoon, Kyong-Ah;Sohn, Ji Yeon;Lee, Eunyoung;Lee, Hyewon;Eom, Hyeon-Seok;Kong, Sun-Young
    • Genomics & Informatics
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    • v.15 no.3
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    • pp.87-97
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    • 2017
  • Multiple myeloma (MM) is a malignant disease caused by an abnormal proliferation of plasma cells, of which the prognostic factors include chromosomal abnormality, ${\beta}$-2 microglobulin, and albumin. Recently, the term chromothripsis has emerged, which is the massive but highly localized chromosomal rearrangement in response to a one-step catastrophic event. Many studies have shown an association of chromothripsis with the prognosis in several cancers; however, few studies have investigated it in MM. Here, we studied the association between chromothripsis-like patterns and treatment resistance or prognosis. First, we analyzed nine MM cell lines (U266, MM.1S, RPMI8226, KMS-11, KMS-12-BM, KMS-12-PE, KMS-28-BM, KMS-28-PE, and NCI-H929) and bone marrow samples of four patients who were diagnosed with MM by next-generation sequencing-based copy number variation analysis. The frequency of the chromothripsis-like pattern was observed in seven cell lines. We analyzed the treatment-induced chromothripsis-like patterns in KMS-12-BM and KMS-12-PE cells. As a result, breakpoints and chromothripsis-like patterns were increased after drug treatment in the relatively resistant KMS-12-BM. We further analyzed the patients' results according to the therapeutic response, which was divided into sensitive and resistant, as suggested by the International Myeloma Working Group. The chromothripsis-like pattern was more frequently observed in the resistant group. In the sensitive group, the frequency of the chromothripsis-like pattern decreased after treatment, whereas the resistant group showed increased chromothripsis-like patterns after the treatment. These results suggest that the chromothripsis-like pattern is associated with treatment response in MM.

Diagnosis and Sequence Analysis of Japanese yam mosaic virus from Yam (Dioscorea opposita) (마(Dioscorea opposita)에 발생한 Japanese yam mosaic virus 진단 및 염기서열 분석)

  • Lee, Joong-Hwan;Son, Chang-Gi;Kwon, Joong-Bae;Nam, Hyo-Hun;Kim, Yeong-Tae;Kim, Mi Kyeong;Lee, Su-Heon
    • Research in Plant Disease
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    • v.22 no.4
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    • pp.289-292
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    • 2016
  • We surveyed the occurrence of Japanese yam mosaic virus (JYMV) on Yam in Gyeongsangbukdo pronvince from 2013 to 2015. The symptoms of JYMV were yellow stripes and chlorosis in yam leaves and the infection rate was ranged from 33.6% to 40.8%. We determined nucleotide sequence encoding the polyprotein of JYMV isolate BRI from yam leaves using next-generation sequencing (NGS) method. The partial nucleotide portion (7,736 nucleotides) of the genomic RNA of the JYMV isolate BRI has been sequenced (accession No. KU309315). The region sequenced includes a single open reading frame (ORF) encoding a polyprotein composed of 2,497 amino acids containing the coat protein (CP) and 3' untranslated region (UTR). The genomic organization of this isolate shows almost the same to that of other members of JYMV. The JYMV isolate BRI showed 77% to 79% nucleotide identity with the Japanese and Chinese strains and isolates. This is the first report of the genome nucleotide sequence of JYMV from Dioscorea opposita in Korea.

Qualitative and Quantitative Analysis for Microbiome Data Matching between Objects (마이크로바이옴 데이터 일치를 위한 물체들 사이의 정량 및 정성적 분석)

  • You, Hee Sang;Ok, Yeon Jeong;Lee, Song Hee;Lee, So Lip;Lee, Young Ju;Lee, Min Ho;Hyun, Sung Hee
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.3
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    • pp.202-213
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    • 2020
  • Although technological advances have allowed the efficient collection of large amounts of microbiome data for microbiological studies, proper analysis tools for such big data are still lacking. Additionally, analyses of microbial communities using poor databases can lead to misleading results. Hence, this study aimed to design an appropriate method for the analysis of big microbial databases. Bacteria were collected from the fingertips and personal belongings (mobile phones and laptop keyboards) of individuals. The genomic DNA was extracted from these bacteria and subjected to next-generation sequencing by targeting the 16S rRNA gene. The accuracy of the bacterial matching percentage between the fingertips and personal belongings was verified using a formula and an environment-related and human-related database. To design appropriate analysis, the bacterial matching accuracy was calculated based on the following three categories: comparison between qualitative and quantitative analysis, comparisons within same-gender participants as well as all participants regardless of gender, and comparison between the use of a human-related bacterial database (hDB) and environment-related bacterial database (eDB). The results showed that qualitative analysis, comparisons within same-gender participants, and the use of hDB provided relatively accurate results. This study provides an analytical method to obtain accurate results when conducting studies involving big microbiological data using human-derived microorganisms.

CNVDAT: A Copy Number Variation Detection and Analysis Tool for Next-generation Sequencing Data (CNVDAT : 차세대 시퀀싱 데이터를 위한 유전체 단위 반복 변이 검출 및 분석 도구)

  • Kang, Inho;Kong, Jinhwa;Shin, JaeMoon;Lee, UnJoo;Yoon, Jeehee
    • Journal of KIISE:Databases
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    • v.41 no.4
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    • pp.249-255
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    • 2014
  • Copy number variations(CNVs) are a recently recognized class of human structural variations and are associated with a variety of human diseases, including cancer. To find important cancer genes, researchers identify novel CNVs in patients with a particular cancer and analyze large amounts of genomic and clinical data. We present a tool called CNVDAT which is able to detect CNVs from NGS data and systematically analyze the genomic and clinical data associated with variations. CNVDAT consists of two modules, CNV Detection Engine and Sequence Analyser. CNV Detection Engine extracts CNVs by using the multi-resolution system of scale-space filtering, enabling the detection of the types and the exact locations of CNVs of all sizes even when the coverage level of read data is low. Sequence Analyser is a user-friendly program to view and compare variation regions between tumor and matched normal samples. It also provides a complete analysis function of refGene and OMIM data and makes it possible to discover CNV-gene-phenotype relationships. CNVDAT source code is freely available from http://dblab.hallym.ac.kr/CNVDAT/.

Prediction of Genomic Relationship Matrices using Single Nucleotide Polymorphisms in Hanwoo (한우의 유전체 표지인자 활용 개체 혈연관계 추정)

  • Lee, Deuk-Hwan;Cho, Chung-Il;Kim, Nae-Soo
    • Journal of Animal Science and Technology
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    • v.52 no.5
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    • pp.357-366
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    • 2010
  • The emergence of next-generation sequencing technologies has lead to application of new computational and statistical methodologies that allow incorporating genetic information from entire genomes of many individuals composing the population. For example, using single-nucleotide polymorphisms (SNP) obtained from whole genome amplification platforms such as the Ilummina BovineSNP50 chip, many researchers are actively engaged in the genetic evaluation of cattle livestock using whole genome relationship analyses. In this study, we estimated the genomic relationship matrix (GRM) and compared it with one computed using a pedigree relationship matrix (PRM) using a population of Hanwoo. This project is a preliminary study that will eventually include future work on genomic selection and prediction. Data used in this study were obtained from 187 blood samples consisting of the progeny of 20 young bulls collected after parentage testing from the Hanwoo improvement center, National Agriculture Cooperative Federation as well as 103 blood samples from the progeny of 12 proven bulls collected from farms around the Kyong-buk area in South Korea. The data set was divided into two cases for analysis. In the first case missing genotypes were included. In the second case missing genotypes were excluded. The effect of missing genotypes on the accuracy of genomic relationship estimation was investigated. Estimation of relationships using genomic information was also carried out chromosome by chromosome for whole genomic SNP markers based on the regression method using allele frequencies across loci. The average correlation coefficient and standard deviation between relationships using pedigree information and chromosomal genomic information using data which was verified using a parentage test andeliminated missing genotypes was $0.81{\pm}0.04$ and their correlation coefficient when using whole genomic information was 0.98, which was higher. Variation in relationships between non-inbred half sibs was $0.22{\pm}0.17$ on chromosomal and $0.22{\pm}0.04$ on whole genomic SNP markers. The variations were larger and unusual values were observed when non-parentage test data were included. So, relationship matrix by genomic information can be useful for genetic evaluation of animal breeding.

Application of Environmental DNA for Monitoring of Freshwater Fish in Korea (환경유전자의 국내 담수어류 모니터링 적용 연구)

  • Kim, Jeong-Hui;Jo, Hyunbin;Chang, Min-Ho;Woo, Seung-Hyun;Cho, Youngho;Yoon, Ju-Duk
    • Korean Journal of Ecology and Environment
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    • v.53 no.1
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    • pp.63-72
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    • 2020
  • In this study, to discuss on the applicability of eDNA as a new method to investigate fish diversity at streams, we applied eDNA at 4 streams (Geum River, Ji Stream, Hwangji Stream, Seomjin River), where endangered species are inhabits, with conventional survey (cast net and kick net). The average (±standard deviation) number of species investigated by eDNA were 19 species (±4.4), and it was relatively higher than average of conventional survey, 10 species (±4.8). Most of case, in this study, eDNA was more efficient than conventional survey. However, there were errors on species identification of Korean endemic species and aliied species from eDNA, and it seems the universal primer (MiFish primer set) is not suitable for them. Furthermore, some of endangered species, caught by conventional method, was not detected by eDNA. As the present universal primer is not suitable for identify the every freshwater fish species in Korea, the complementing or development of universal primer is needed, and the eDNA application after species specific marker development for detecting specific species like endangered species should be considered. In conclusion, if the manual for field survey method by eDNA is developed, we expect applicability enlargement for water ecosystem survey.

Enrichment and verification of differentially expressed miRNAs in bursa of Fabricius in two breeds of duck

  • Luo, Jun;Liu, Junying;Liu, Hehe;Zhang, Tao;Wang, Jiwen;He, Hua;Han, Chunchun
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.7
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    • pp.920-929
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    • 2017
  • Objective: The bursa of Fabricius (BF) is a central humoral immune organ belonging specifically to avians. Recent studies had suggested that miRNAs were active regulators involved in the immune processes. This study was to investigate the possible differences of the BF at miRNA level between two genetically disparate duck breeds. Methods: Using Illumina next-generation sequencing, the miRNAs libraries of ducks were established. Results: The results showed that there were 66 differentially expressed miRNAs and 28 novel miRNAs in bursa. A set of abundant miRNAs (i.e., let-7, miR-146a-5p, miR-21-5p, miR-17~92) which are involved in immunity and disease were detected and the predicted target genes of the novel miRNAs were associated with duck high anti-adversity ability. By gene ontology analysis and enriching KEGG pathway, the targets of differential expressed miRNAs were mainly involved in immunity and disease, supporting that there were differences in the BF immune functions between the two duck breeds. In addition, the metabolic pathway had the maximum enriched target genes and some enriched pathways that were related to cell cycle, protein synthesis, cell proliferation and apoptosis. It indicted that the difference of metabolism may be one of the reasons leading the immune difference between the BF of two duck breeds. Conclusion: This data lists the main differences in the BF at miRNAs level between two genetically disparate duck breeds and lays a foundation to carry out molecular assisted breeding of poultry in the future.

Transcriptome profiling and comparative analysis of Panax ginseng adventitious roots

  • Jayakodi, Murukarthick;Lee, Sang-Choon;Park, Hyun-Seung;Jang, Woojong;Lee, Yun Sun;Choi, Beom-Soon;Nah, Gyoung Ju;Kim, Do-Soon;Natesan, Senthil;Sun, Chao;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • v.38 no.4
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    • pp.278-288
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    • 2014
  • Background: Panax ginseng Meyer is a traditional medicinal plant famous for its strong therapeutic effects and serves as an important herbal medicine. To understand and manipulate genes involved in secondary metabolic pathways including ginsenosides, transcriptome profiling of P. ginseng is essential. Methods: RNA-seq analysis of adventitious roots of two P. ginseng cultivars, Chunpoong (CP) and Cheongsun (CS), was performed using the Illumina HiSeq platform. After transcripts were assembled, expression profiling was performed. Results: Assemblies were generated from ~85 million and ~77 million high-quality reads from CP and CS cultivars, respectively. A total of 35,527 and 27,716 transcripts were obtained from the CP and CS assemblies, respectively. Annotation of the transcriptomes showed that approximately 90% of the transcripts had significant matches in public databases.We identified several candidate genes involved in ginsenoside biosynthesis. In addition, a large number of transcripts (17%) with different gene ontology designations were uniquely detected in adventitious roots compared to normal ginseng roots. Conclusion: This study will provide a comprehensive insight into the transcriptome of ginseng adventitious roots, and a way for successful transcriptome analysis and profiling of resource plants with less genomic information. The transcriptome profiling data generated in this study are available in our newly created adventitious root transcriptome database (http://im-crop.snu.ac.kr/transdb/index.php) for public use.