• Title/Summary/Keyword: Expressed sequence tags (ESTs)

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Functional Analysis of ESTs from the 14-year Root of Korean Ginseng

  • Yang, Deok-Chun;In, Jun-Gyo;Kim, Moo-Sung;Jeon, Jong-Seong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.125-125
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    • 2003
  • To assist genetic study of the root development in Panax ginseng, which is one of the most important medicinal plant, expressed sequence tags (EST) analysis was carried out. We constructed a cDNA library using the 14-year ginseng root. Partial sequences were obtained from 2,975 clone. The ESTs could be clustered into 1,991 (70.2%) non-redundant groups. Similarity search of the non-redundant ESTs against public non-redundant databases of both protein and DNA indicated that 1,553 groups show similarity to genes of blown function. These ESTs clones were divided into sixteen categories depending upon gene function. The most abundant transcripts were ribonuclease 1 (67) and ribonuclease 2 (65). Our extensive EST analysis of genes expressed in 14-year ginseng root not only contributes to the understanding of the dynamics of genome expression patterns in root organ but also adds data to the reperoire of all genomic genes.

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Cataloguing of Anther Expressed Genes through Differential Slot Blot in Oriental Lily (Lilium Oriental Hybrid 'Acapulco') (아카풀코나리에서 Differential Slot Blot을 이용한 약발현 유전자 목록작성)

  • Suh, Eun-Jung;Yu, Hee Ju;Han, Bong Hee;Lim, Yong Pyo;Jeong, Mi-Jeong;Lee, Seong-Kon;Kim, Dong-Hern;Chang, An-Cheol;Yae, Byeong Woo
    • Horticultural Science & Technology
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    • v.31 no.5
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    • pp.598-606
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    • 2013
  • Anther is the major organ of flower in responsible to reproduction and outward appearance. From anther-specific cDNA library of Lilium Oriental Hybrid 'Acapulco', 2000 expressed sequence tags were selected randomly. Differential slot blot analysis with cDNA probes from the anther and leaf was used to get anther-expressed clone and 570 non-redundant ESTs were obtained and sequenced. Compared to the GenBank database using BLASTX algorithm, 191 clones showed significant similarity but others (66.5%) did not measured to known sequence. Functional categories according to gene ontology (GO) annotation included sequence representing a significant portion of protein in cell and cell part respectively. A transcriptional analysis at 7 different organs and developmental stage was performed using northern blot with thirty ESTs as putative anther specific gene. This report suggest that selection of anther expressed clone using differential slot blot was considered as very effective tool and our current study can provide fundamental information on the lily anther including pollen furthermore.

Current Status of Comparative Mapping in Livestock

  • Lee, J.H.;Moran, C.;Park, C.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.10
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    • pp.1411-1420
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    • 2003
  • Comparative maps, representing chromosomal locations of homologous genes in different species, are useful sources of information for identifying candidate disease genes and genes determining complex traits. They facilitate gene mapping and linkage prediction in other species, and provide information on genome organization and evolution. Here, the current gene mapping and comparative mapping status of the major livestock species are presented. Two techniques were widely used in comparative mapping: FISH (Fluorescence In Situ Hybridization) and PCR-based mapping using somatic cell hybrid (SCH) or radiation hybrid (RH) panels. New techniques, using, for example, ESTs (Expressed Sequence Tags) or CASTS (Comparatively Anchored Sequence Tagged Sites), also have been developed as useful tools for analyzing comparative genome organization in livestock species, further enabling accurate transfer of valuable information from one species to another.

Detection and Prediction of Alternative Splicing with One-leaf One-node Tree (One-leaf One-node 트리를 이용한 선택 스플라이싱 탐지 및 예측)

  • Park, Min-Seo
    • The Journal of the Korea Contents Association
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    • v.10 no.10
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    • pp.102-110
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    • 2010
  • Alternative splicing is an important process in gene expression. Alternative Splicing can lead to mutations and diseases. Most studies detect alternatively spliced genes with ESTs (Expressed Sequence Tags). However, reliance on ESTs might have some weaknesses in predicting alternative splicing. ESTs have been stored in the libraries. The EST libraries are often not clearly organized and annotated. We can pick erroneous ESTs. It is also difficult to predict whether or not alternative splicing exists for those genes where ESTs are not available. To address these issues and to improve the quality of detection and prediction for alternative splicing, we propose the One-leaf One-node Tree Algorithm that uses pre-mRNAs. It is achieved by codons, three nucleotides, as attributes for each chromosome in Arabidopsis thaliana. The proposed decision tree shows that alternative and normal splicing have different splicing patterns according to triplet nucleotides in each chromosome. Based on the patterns, alternative splicing of unlabeled genes can also be predicted.

Construction of a full-length cDNA library from Pinus koraiensis and analysis of EST dataset (잣나무(Pinus koraiensis)의 cDNA library 제작 및 EST 분석)

  • Kim, Joon-Ki;Im, Su-Bin;Choi, Sun-Hee;Lee, Jong-Suk;Roh, Mark S.;Lim, Yong-Pyo
    • Korean Journal of Agricultural Science
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    • v.38 no.1
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    • pp.11-16
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    • 2011
  • In this study, we report the generation and analysis of a total of 1,211 expressed sequence tags (ESTs) from Pinus koraiensis. A cDNA library was generated from the young leaf tissue and a total of 1,211 cDNA were partially sequenced. EST and unigene sequence quality were determined by computational filtering, manual review, and BLAST analyses. In all, 857 ESTs were acquired after the removal of the vector sequence and filtering over a minimum length 50 nucleotides. A total of 411 unigene, consisting of 89 contigs and 322 singletons, was identified after assembling. Also, we identified 77 new microsatellite-containing sequences from the unigenes and classified the structure according to their repeat unit. According to homology search with BLASTX against the NCBI database, 63.1% of ESTs were homologous with known function and 22.2% of ESTs were matched with putative or unknown function. The remaining 14.6% of ESTs showed no significant similarity to any protein sequences found in the public database. Gene ontology (GO) classification showed that the most abundant GO terms were transport, nucleotide binding, plastid, in terms biological process, molecular function and cellular component, respectively. The sequence data will be used to characterize potential roles of new genes in Pinus and provided for the useful tools as a genetic resource.

Mining of Biomarker Genes from Expressed Sequence Tags and Differential Display Reverse Transcriptase-Polymerase Chain Reaction in the Self-fertilizing Fish, Kryptolebias marmoratus and Their Expression Patterns in Response to Exposure to an Endocrine-disrupting Alkylphenol, Bisphenol A

  • Lee, Young-Mi;Rhee, Jae-Sung;Hwang, Dae-Sik;Kim, Il-Chan;Raisuddin, Sheikh;Lee, Jae-Seong
    • Molecules and Cells
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    • v.23 no.3
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    • pp.287-303
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    • 2007
  • Expressed sequence tags (ESTs) and differentially expressed cDNAs from the self-fertilizing fish, Kryptolebias marmoratus were mined to develop alternative biomarkers for endocrine-disrupting chemicals (EDCs). 1,577 K. marmoratus cDNA clones were randomly sequenced from the 5'-end. These clones corresponded to 1,518 and 1,519 genes in medaka dbEST and zebrafish dbEST, respectively. Of the matched genes, 197 and 115 genes obtained Unigene IDs in medaka dbEST and zebrafish dbEST, respectively. Many of the annotated genes are potential biomarkers for environmental stresses. In a differential display reverse transcriptase-polymerase chain reaction (DD RT-PCR) study, 56 differential expressed genes were obtained from fish liver exposed to bisphenol A. Of these, 16 genes were identified after BLAST search to GenBank, and the annotated genes were mainly involved in catalytic activity and binding. The expression patterns of these 16 genes were validated by real-time RT-PCR of liver tissue from fish exposed to bisphenol A. Our findings suggest that expression of these 16 genes is modulated by endocrine disrupting chemicals, and therefore that they are potential biomarkers for environmental stress including EDCs exposure.

The EST Analysis and Transgene Expression System in Rice

  • Kim, Jukon;Nahm, Baek-Hie
    • Journal of Plant Biotechnology
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    • v.1 no.1
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    • pp.46-55
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    • 1999
  • The expressed sequence tags(ESTs) from immature seed of rice, Oryza sativa cv Milyang 23, were partially sequenced and analyzed by homology. As of 1998, the partial sequences of about 6,600 cDNA clones were analyzed from normal and normalized immature seed cDNA libraries. About 2,200 ESTs were putatively identified by BLASTX deduced amino acid sequence homology analysis. About 20% of them were putatively identified as storage proteins. Also the clones were highly homologous to genes involved particularly in starch biosynthesis, glycolysis, signal transduction and defenses. Compared to 35% of redundancy in the ESTs of normal cDNA library, that from the substracted library was 15%. The Korea Rice Genome Network is maintained to provide the updated information of sequences, their homologies and sequence alignments of ESTs. For the stable expression of transgene in rice, diverse vectors were developed for overexpression, targeting and gene dosage effect with transit peptides (Tp) and matrix attachment region (MAR) sequence from chicken lysozyme locus. The rice calli were transformed via Agrobacterium tumefaciens LBA4404(pSB1) with the triparental mating technique and selected by herbicide resistance. The green fluorescent protein(GFP) gene in expression vector under the control of rbcS promoter-Tp was overexpressed upto 10 % of the total soluble protein. In addition, the Tp-sGFP fusion protein was properly processed during translocation into chloroplast. The expression of sGFP in the presence of MAR sequences was analyzed with Northern and immunoblot analysis. All the lines in which sGFP transgene with MAR sequence, showed position independent and copy number-dependent expression, while the lines without MAR showed the varied level of expression with the integration site. Thus the MAR sequence significantly reduced the variation in transgene expression between independent transformants.

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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.

Gene Expression Profiling of Eukaryotic Microalga, Haematococcus pluvialis

  • EOM HYUNSUK;PARK SEUNGHYE;LEE CHOUL-GYUN;JIN EONSEON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1060-1066
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    • 2005
  • Under environmental stress, such as strong irradiance or nitrogen deficiency, unicellular green algae of the genus Haematococcus accumulate secondary carotenoids, i.e. astaxanthin, in the cytosol. The induction and regulation of astaxanthin biosynthesis in microalgae has recently received considerable attention owing to the increasing use of secondary carotenoids as a source of pigmentation for fish aquacultures, and as a potential drug in cancer prevention as a free-radical quencher. Accordingly, this study generated expressed sequence tags (ESTs) from a library constructed from astaxanthin-induced Haematococcus pluvialis. Partial sequences were obtained from the 5' ends of 1,858 individual cDNAs, and then grouped into 1,025 non-overlapping sequences, among which 708 sequences were singletons, while the remainder fell into 317 clusters. Approximately $63\%$ of the EST sequences showed similarity to previously described sequences in public databases. H. pluvialis was found to consist of a relatively high percentage of genes involved in genetic information processing ($15\%$) and metabolism ($11\%$), whereas a relatively low percentage of sequences was involved in the signal transduction ($3\%$), structure ($2\%$), and environmental information process ($3\%$). In addition, a relatively large fraction of H. pluvialis sequences was classified as genes involved in photosynthesis ($9\%$) and cellular process ($9\%$). Based on this EST analysis, the full-length cDNA sequence for superoxide dismutase (SOD) of H. pluvialis was cloned, and the expression of this gene was investigated. The abundance of SOD changed substantially in response to different culture conditions, indicating the possible regulation of this gene in H. pluvialis.