• Title/Summary/Keyword: EST (Expressed Sequence Tag)

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Expressed Sequence Tags in Rainbow Trout (Oncorhynchus mykiss) Kidney and Microarray Analysis in Young and Old Kidney (무지개송어 신장으로부터 EST 발굴 및 연령에 따른 유전자 발현 분석)

  • Kim, Soon-Hag;Shin, Yong-Kook;Bang, In-Chul
    • Journal of Life Science
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    • v.13 no.1
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    • pp.128-135
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    • 2003
  • 102 ESTs (Expressed Sequence Tags) were obtained by sequencing clones from a library of rainbow trout kidney cDNAs. Of the sequences generated, 55.8% of the ESTs were represented by 37 known genes. The 45 clones of unknown gene products potentially represent 40 novel genes. The genes involved in structural function (14.5%) and transcription/translation (11.6%) account for the major gene expression activities in the kidney Microarray experiment was conducted to compare gene expression of the unique ESTs in young and adult rainbow trout kidneys. While mitochondrion, cytochrome b, rho G, spastin protein, and three unknown genes were down-regulated in the mature fish kidney, calponin 1, calcium binding protein, histone deacetylase 1, and an unknown gene were up-regulated in the mature fish kidney. This research demonstrates the feasibility and power of functional genomics in rainbow trout.

Bioinformatics Approach to Direct Target Prediction for RNAi Function and Non-specific Cosuppression in Caenorhabditis elegans (생물정보학적 접근을 통한 Caenorhabditis elegans 모델시스템의 생체내 RNAi 기능예측 및 비특이적 공동발현억제 현상 분석)

  • Kim, Tae-Ho;Kim, Eui-Yong;Joo, Hyun
    • KSBB Journal
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    • v.26 no.2
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    • pp.131-138
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    • 2011
  • Some computational approaches are needed for clarifying RNAi sequences, because it takes much time and endeavor that almost of RNAi sequences are verified by experimental data. Incorrectness of RNAi mechanism and other unaware factors in organism system are frequently faced with questions regarding potential use of RNAi as therapeutic applications. Our massive parallelized pair alignment scoring between dsRNA in Genebank and expressed sequence tags (ESTs) in Caenorhabditis elegans Genome Sequencing Projects revealed that this provides a useful tool for the prediction of RNAi induced cosuppression details for practical use. This pair alignment scoring method using high performance computing exhibited some possibility that numerous unwanted gene silencing and cosuppression exist even at high matching scores each other. The classifying the relative higher matching score of them based on GO (Gene Ontology) system could present mapping dsRNA of C. elegans and functional roles in an applied system. Our prediction also exhibited that more than 78% of the predicted co-suppressible genes are located in the ribosomal spot of C. elegans.

Liuux Cluster based Biological Sequence Parallel Processing Model Development and Efficiency Verification (리눅스 클러스터기반 유전자서열분석 병렬처리 모형 개발 및 성능 검증)

  • 박미화;김재우;박춘규;유승식
    • Proceedings of the Korean Information Science Society Conference
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    • 2003.04a
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    • pp.106-108
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    • 2003
  • Human Genome Project와 같은 대형 Sequencing 프로젝트와 High-throughput Sequencing 기술의 발전으로 현재 Expressed Sequence Tag (EST)와 같은 대량의 DNA 서열들이 생산되고 있다. 이를 효과적이고 효율적으로 분석해야 할 필요성이 증대되고 있다. 대부분의 실험자들이 서열 분석을 위해 우선적으로 BLAST 검색을 이용하고 있다. 하지만 대량의 서열, 검색 DB의 크기, BLAST 검색 결과의 복잡성에 의해 어려움을 겪고 있다. 이에 빠르고 정리된 결과를 보여줄 수 있는 BLAST 검색 시스템의 필요성이 커지고 있다. 이에 본 논문은 미국 생명공학연구소(NCBI)에서 제공하는 유전자 서열 검색 툴인 BLAST(Basic Logical Alignment Tool)를 클러스터 수퍼 컴퓨터 구축 기술을 기반으로 한 병렬처리와 Gene Ontology를 이용하여 방대한 양의 서열 검색 결과를 요약하는 모형을 제시한다. 이것은 신약개발 및 유전자 발굴 등의 연구기간을 획기적으로 단축시켜 신약 개 발, 농업, 화학, 의료, 환경 등 생명공학 연구에 핵심적인 역할을 할 수 있다. 또한 성능 실험을 통하여 분석결과 대기시간을 최소화하는 병렬처리모형의 효율성을 검증하였다.

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EST Knowledge Integrated Systems (EKIS): An Integrated Database of EST Information for Research Application

  • Kim, Dae-Won;Jung, Tae-Sung;Choi, Young-Sang;Nam, Seong-Hyeuk;Kwon, Hyuk-Ryul;Kim, Dong-Wook;Choi, Han-Suk;Choi, Sang-Heang;Park, Hong-Seog
    • Genomics & Informatics
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    • v.7 no.1
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    • pp.38-40
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    • 2009
  • The EST Knowledge Integrated System, EKIS (http://ekis.kribb.re.kr), was established as a part of Korea's Ministry of Education, Science and Technology initiative for genome sequencing and application research of the biological model organisms (GEAR) project. The goals of the EKIS are to collect EST information from GEAR projects and make an integrated database to provide transcriptomic and metabolomic information for biological scientists. The EKIS constitutes five independent categories and several retrieval systems in each category for incorporating massive EST data from high-throughput sequencing of 65 different species. Through the EKIS database, scientists can freely access information including BLAST functional annotation as well as Genechip and pathway information for KEGG. By integrating complex data into a framework of existing EST knowledge information, the EKIS provides new insights into specialized metabolic pathway information for an applied industrial material.

Construction and Characterization of a cDNA Library from the Camelina sativa L. as an Alternative Oil-Seed Crop (신 바이오디젤 원료 작물인 Camelina의 cDNA library 제작 및 유전자 특성)

  • Park, Won;Jang, Young-Seok;Ahn, Sung-Ju
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.55 no.2
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    • pp.151-158
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    • 2010
  • Camelina sativa L., known as popular names "gold-of-pleasure" or "false flax" is an alternative oilseed crop that can be grown under different climatic and soil conditions. Up to date, however, the genomic information of Camelina has not been studied in detail. Therefore, a cDNA library was constructed and characterized from young leaves. The constructed cDNA library incorporated of 1334 cDNA clones and the size of the insertion fragments average was 736 base pair. We generated a total of 1269 high-quality expressed sequence tags (ESTs) sequences. The result of cluster analysis of EST sequences showed that the number of unigene was 851. According to subsequent analysis, the 476 (55.9%) unigenes were highly homologous to known function genes and the other 375 (44.1%) unigenes were unknown. Remaining 63 (7.4%) unigenes had no homology with any other peptide in NCBI database, indicating that these seemed to be novel genes expressed in leaves of Camelina. The database-matched ESTs were further classified into 17 categories according to their functional annotation. The most abundant of categories were "protein with binding function or cofactor requirement (27%)", "metabolism (11%)", "subcellular localization (11%)", "cellular transport, transport facilities and transport routes (7%)", "energy (6%)", "regulation of metabolism and protein function (6%)". Our result in this study provides an overview of mRNA expression profile and a basal genetic information of Camelina as an oilseed crop.

Analysis of Genetic Polymorphism and Relationship of Korean Ginseng Cultivars and Breeding Lines using EST-SSR Marker (EST-SSR 마커를 이용한 인삼 품종과 육성계통의 유전적 다형성 및 유연관계 분석)

  • Bang, Kyong-Hwan;Seo, A-Yeon;Chung, Jong-Wook;Kim, Young-Chang;Jo, Ick-Hyun;Kim, Jang-Uk;Kim, Dong-Hwi;Cha, Seon-Woo;Cho, Yong-Gu;Kim, Hong-Sig
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.4
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    • pp.277-285
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    • 2012
  • In this study, Expressed Sequence Tag-Simple Sequence Repeat (EST-SSR) analyses were used to clarify the genetic polymorphisms among Korean ginseng cultivars and breeding lines and to classify them into distinct genetic groups. Polymorphic and reproducible bands were produced by 14 primers out of total 30 primers used in this study. Fourteen EST-SSR loci generated a total of 123 bands. Amplified PCR products showed the highly reproducible banding patterns at 110~920 bp. The number of amplified bands for each EST-SSR primers ranged from 2 to 19 with a mean of 8.8 bands. P26 and P35 primers showed 13 and 12 banding patterns, respectively. The number of alleles for each EST-SSR locus ranged from 1.67 to 2.00 with a mean of 1.878 alleles. P34 and P60 primers showed the highest and the lowest genetic polymorphism, respectively. Cluster analysis based on genetic similarity estimated by EST-SSR markers classified Korean cultivars and breeding lines into 4 groups. Group included Gopoong and Chunpoong and 9 breeding lines (55%), group included 2 breeding lines (10%), group included 3 breeding lines (15%), group included Gumpoong and 3 breeding lines (20%). Consequently, the EST-SSR marker developed in this study may prove useful for the evaluation of genetic diversity and differentiation of Korean ginseng cultivars and breeding lines.

Analysis of Expressed Sequence Tags from the Red Alga Griffithsia okiensis

  • Lee, Hyoung-Seok;Lee, Hong-Kum;An, Gyn-Heung;Lee, Yoo-Kyung
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.541-546
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    • 2007
  • Red algae are distributed globally, and the group contains several commercially important species. Griffithsia okiensis is one of the most extensively studied red algal species. In this study, we conducted expressed sequence tag (ESTs) analysis and synonymous codon usage analysis using cultured G. okiensis samples. A total of 1,104 cDNA clones were sequenced using a cDNA library made from samples collected from Dolsan Island, on the southern coast of Korea. The clustering analysis of these sequences allowed for the identification of 1,048 unigene clusters consisting of 36 consensus and 1,012 singleton sequences. BLASTX searches generated 532 significant hits (E-value <$10^{-4}$) and via further Gene Ontology analysis, we constructed a functional classification of 434 unigenes. Our codon usage analysis showed that unigene clusters with more than three ESTs had higher GC contents (76.5%) at the third position of the codons than the singletons. Also, the majority of the optimal codons of G. okiensis and Chondrus crispus belonging to Bangiophycidae were G-ending, whereas those of Porphyra yezoensis belonging to Florideophycidae were G-ending. An orthologous gene search for the P. yezoensis EST database resulted in the identification of 39 unigenes commonly expressed in two rhodophytes, which have putative functions for structural proteins, protein degradation, signal transduction, stress response, and physiological processes. Although experiments have been conducted on a limited scale, this study provides a material basis for the development of microarrays useful for gene expression studies, as well as useful information for the comparative genomic analysis of red algae.

Evaluation of horticultural traits and genetic relationship in melon germplasm (멜론 유전자원의 원예형질 특성 및 유연관계 분석)

  • Jung, Jaemin;Choi, Sunghwan;Oh, Juyeol;Kim, Nahui;Kim, Daeun;Son, Beunggu;Park, Younghoon
    • Journal of Plant Biotechnology
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    • v.42 no.4
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    • pp.401-408
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    • 2015
  • Horticultural traits and genetic relationship were evaluated for 83 melon (Cucumis melo L.) cultivars. Survey of a total of 36 characteristics for seedling, leaf, stem, flower, fruit, and seed and subsequent multiple analysis of variance (MANOVA) were conducted. Principal component analysis (PCA) showed that 8 principle components including fruit weight, fruit length, fruit diameter, cotyledon length, seed diameter, and seed length accounted for 76.3% of the total variance. Cluster analysis of the 83 melon cultivars using average linkage method resulted in 5 clusters at coefficient of 0.7. Cluster I consisted of cultivars with high values for fruit-related traits, Cluster II for soluble solid content, and Cluster V for high ripening rate. Genotyping of the 83 cultivars was conducted using 15 expressed-sequence tagged-simple sequence repeat (EST-SSR) from the Cucurbit Genomics Initiative (ICuGI) database. Analysis of genetic relatedness by UPGMA resulted in 6 clusters. Mantel test indicated that correlation between morphological and genetic distance was very low (r = -0.11).

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.

In silico analysis of candidate genes involved in light sensing and signal transduction pathways in soybean

  • Quecini, V.;Zucchi, M.I.;Pinheiro, J.B.;Vello, N.A.
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.59-73
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    • 2008
  • Several aspects of photoperception and light signal transduction have been elucidated by studies with model plants. However, the information available for economically important crops, such as Fabaceae species, is scarce. In order to incorporate the existing genomic tools into a strategy to advance soybean research, we have investigated publicly available expressed sequence tag (EST) sequence databases in order to identify Glycine max sequences related to genes involved in light-regulated developmental control in model plants. Approximately 38,000 sequences from open-access databases were investigated, and all bona fide and putative photoreceptor gene families were found in soybean sequence databases. We have identified G. max orthologs for several families of transcriptional regulators and cytoplasmic proteins mediating photoreceptor-induced responses, although some important Arabidopsis phytochrome-signaling components are absent. Moreover, soybean and Arabidopsis genefamily homologs appear to have undergone a distinct expansion process in some cases. We propose a working model of light perception, signal transduction and response-eliciting in G. max, based on the identified key components from Arabidopsis. These results demonstrate the power of comparative genomics between model systems and crop species to elucidate several aspects of plant physiology and metabolism.