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

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Survey of Expressed Sequence Tags from Tissue-Specific cDNA Libraries in Hemibarbus mylodon, an Endangered Fish Species (멸종위기 어류 어름치 Hemibarbus mylodon (Cypriniformes)로부터 조직별 EST library 제작 및 발현 유전자 탐색)

  • Bang, In-Chul;Lim, Yoon-Hee;Cho, Young-Sun;Lee, Sang-Yoon;Nam, Yoon-Kwon
    • Journal of Aquaculture
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    • v.20 no.4
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    • pp.248-254
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    • 2007
  • Representative cDNA libraries were constructed from various tissue sources of Hemibarbus mylodon, an endangered freshwater fish species in Korea, for the mining of expressed sequence tags (ESTs). Randomized and non-normalized EST analysis was performed with 7 unidirectional cDNA libraries generated from brain, intestine, kidney, liver, muscle, ovary or testis. Of 3,383 ESTs in total, the number of singleton was 2,029, and 333 contigs containing 1,354 ESTs were assembled (percent of unigene = 70.0%). Abundantly expressed gene transcripts and broad clustering of putative gene function were tissue-specific in general, and the redundancy was also variable among those libraries. Over half of H. mylodon ESTs were matched with orthologues from other teleosts among which zebrafish gene sequences were the most frequent in those matches. This initial setting of EST libraries achieved in the present study would be a fundamental basis for the banking of gene resources from this endangered fish species.

Expressed Sequence Tag Analysis of Olive Flounder (Paralichthys olivaceus): Genes and Expression Profile from the Liver

  • Lee Jeong-Ho;Kim Young-Ok;Kim Kyung-Kil;Kim Woo-Jin;Park Doo-Won;Park Jung-Youn;Kim Kyu-Won
    • Fisheries and Aquatic Sciences
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    • v.6 no.3
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    • pp.110-115
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    • 2003
  • Expressed sequence tag (EST) analysis was conducted using a cDNA library made from the liver mRNA of olive flounder (Paralichthys olivaceus). In the survey of 421 ESTs, 362 showed significant homology to previously described genes while 59 were unidentified or novel. Comparative analysis of the identified ESTs showed that 69 $(19.0\%)$ clones were identified as homologous to the previously reported olive flounder ESTs, and 279 $(77.1\%)$ clones were identified as orthologs of known genes from other organisms. The remaining 14 $(3.9\%)$ clones were identified as orthologs of known sequences with unknown functions. These tagged cDNA clones, identified and unidentified, could provide fundamental baseline data for genomic studies of this species.

Expressed sequence tags analysis of Blattella germanica

  • CHUNG Hyang Suk;YU Tai Hyun;KIM Bong Jin;KIM Sun Mi;KIM Joo Yeong;YU Hak Sun;Jeong Hae Jin;OCK Mee Sun
    • Parasites, Hosts and Diseases
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    • v.43 no.4 s.136
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    • pp.149-156
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    • 2005
  • Four hundred and sixty five randomly selected clones from a cDNA library of Blattella germanica were partially sequenced and searched using BLAST as a means of analyzing the transcribed sequences of its genome. A total of 363 expressed sequence tags (ESTs) were generated from 465 clones after editing and trimming the vector and ambiguous sequences. About $42\%$ (154/363) of these clones showed significant homology with other data base registered genes. These new B. germanica genes constituted a broad range of transcripts distributed among ribosomal proteins, energy metabolism, allergens, proteases, protease inhibitors, enzymes, translation, cell signaling path-ways, and proteins of unknown function. Eighty clones were not well-matched by database searches, and these rep-resent new B. germanica-specific ESTs. Some genes which drew our attention are discussed. The information obtained increases our understanding of the B. germanica genome.

Analysis of Expressed Sequence Tags from the Wood-Decaying Fungus Fomitopsis palustris and Identification of Potential Genes Involved in the Decay Process

  • Karim, Nurul;Shibuya, Hajime;Kikuchi, Taisei
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.347-358
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    • 2011
  • Fomitopsis palustris, a brown-rot basidiomycete, causes the most destructive type of decay in wooden structures. In spite of its great economic importance, very little information is available at the molecular level regarding its complex decay process. To address this, we generated over 3,000 expressed sequence tags (ESTs) from a cDNA library constructed from F. palustris. Clustering of 3,095 high-quality ESTs resulted in a set of 1,403 putative unigenes comprising 485 contigs and 918 singlets. Homology searches based on BlastX analysis revealed that 78% of the F. palustris unigenes had a significant match to proteins deposited in the nonredundant databases. A subset of F. palustris unigenes showed similarity to the carbohydrateactive enzymes (CAZymes), including a range of glycosyl hydrolase (GH) family proteins. Some of these CAZyme-encoded genes were previously undescribed for F. palustris but predicted to have potential roles in biodegradation of wood. Among them, we identified and characterized a gene (FpCel45A) encoding the GH family 45 endoglucanase. Moreover, we also provided functional classification of 473 (34%) of F. palustris unigenes using the Gene Ontology hierarchy. The annotated EST data sets and related analysis may be useful in providing an initial insight into the genetic background of F. palustris.

Analyses of Expressed Sequence Tags from Chironomus riparius Using Pyrosequencing : Molecular Ecotoxicology Perspective

  • Nair, Prakash M. Gopalakrishnan;Park, Sun-Young;Choi, Jin-Hee
    • Environmental Analysis Health and Toxicology
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    • v.26
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    • pp.10.1-10.7
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    • 2011
  • Objects: Chironomus riparius, a non-biting midge (Chironomidae, Diptera), is extensively used as a model organism in aquatic ecotoxicological studies, and considering the potential of C. riparius larvae as a bio-monitoring species, little is known about its genome sequences. This study reports the results of an Expressed Sequence Tags (ESTs) sequencing project conducted on C. riparius larvae using 454 pyrosequencing. Method: To gain a better understanding of C. riparius transcriptome, we generated ESTs database of C.ripairus using pyrosequencing method. Results: Sequencing runs, using normalized cDNA collections from fourth instar larvae, yielded 20,020 expressed sequence tags, which were assembled into 8,565 contigs and 11,455 singletons. Sequence analysis was performed by BlastX search against the National Center for Biotechnology Information (NCBI) nucleotide (nr) and uniprot protein database. Based on the gene ontology classifications, 24% (E-value${\leq}1^{-5}$) of the sequences had known gene functions, 24% had unknown functions and 52% of sequences did not match any known sequences in the existing database. Sequence comparison revealed 81% of the genes have homologous genes among other insects belonging to the order Diptera providing tools for comparative genome analyses. Targeted searches using these annotations identified genes associated with essential metabolic pathways, signaling pathways, detoxification of toxic metabolites and stress response genes of ecotoxicological interest. Conclusions: The results obtained from this study would eventually make ecotoxicogenomics possible in a truly environmentally relevant species, such as, C. riparius.

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.

Characterization of Expressed Sequence Tags (ESTs) Generated from the Bombyx mandarina Whole Larvae and Molecular Cloning of Serine Protease Homologue Gene

  • Hwang, Jae Sam;Yun, Eun Young;Goo, Tae Won;Kim, Iksoo;Choi, Kwang Ho;Seong, Su Il;Kim, Keun Young;Lee, Sang Mong;Kang, Seok Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.2
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    • pp.167-171
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    • 2004
  • We constructed an oligo-d(T) primed directional cDNA library from the Bombyx mandarina whole larvae. In an effort to isolate genes expressed in the B. mandarina, 227 expressed sequence tags (ESTs) were generated by single-pass sequencing from the cDNA library. Sequence analysis showed that 107 clones (47.1%) were classified into known genes and 120 clones (52.9%) were novel transcripts, which are unknown for their function. Of the 107 known genes, the most abundant gene was found to be actin and followed by serine protease in the expression profile. Among these clones, a serine protease homolog (BmSP) which is a class of proteolytic enzymes isolated. Full-length sequence of the BmSP cDNA clone was 922 bp in length and has an open reading frame of 276 amino acids. The conserved histidine, aspatic acid and serine residues forming the catalytic center as well as cysteine residues contributing to three disulphide bonds also were found in Bmsp gene. mRNA expression analysis revealed a high and specific expression of the gene only in midgut tissue, suggesting that BmSP gene is closely associated with the expression of digestive enzyme.

Analysis of Expressed Sequence Tags generated from Uroctea lesserti Schenkel(Araneal)

  • Park, Kwang-Ho;Hong, Sun-Mee;Hwang, Jae-Sam;Goo, Tae-Won;Yun, Eun-Young;Kim, Nam-Soon;Kang, Seok-Woo
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.141-142
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    • 2003
  • Partial cDNA sequencing to generate expressed sequence tags (ESTs) is being used at present for the fast and efficient obtainment of a detailed profile of genes expressed in various tissues, cell types, or developmental stages. We describe here the construction, DNA sequencing and sequence profiles of cDNA library from Uroctea lesserti. (omitted)

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Expressed Sequence Tags of the Wheat-rye Translocation Line Possessing 2BS/2RL

  • Jang, Cheol-Seong;Hong, Byung-Hee;Seo, Yong-Weon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.3
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    • pp.302-307
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    • 1999
  • Hamlet (PI549276) possessing 2RL was obtained by cross between a wheat cultivar ND7532 (Froid/Centurk) and a rye cultivar Chaupon. Chaupon was known to have resistant gene to biotype L of Hessian fly [Mayetiola destructor (Say)] larvae. The wheat-rye translocation line (Coker797*4/Hamlet) was also known to be resistant to biotype L of Hessian fly larvae. We analysed a set of 96 ESTs from the wheat-rye translocation line (2BS/2RL). ESTs were classified by various physiological processings, such as primary metabolism, secondary metabolism, transcription, translation, transport, signal transduction, defense, transposable element, and others. Three sequences encoding thioredoxin peroxidase, 26S rRNA, and rubisco small subunits were homologous to registered genes in rye. Although limited number of clones were used to develop ESTs, these clones and their sequence information may be useful for researchers studying general physiology and molecular biology on the translocation line.

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