• Title/Summary/Keyword: Expressed Sequence Tags

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Identification of a Novel Gene by EST Clustering and its Expression in Mouse Ovary and Testis (EST Clustering 방법으로 동정한 새로운 유전자의 생쥐 난소 및 정소에서의 발현)

  • Hwang, Sang-Joon;Park, Chang-Eun;Hwang, Kyu-Chan;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.4
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    • pp.253-263
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    • 2006
  • Objective: Identification of the regulatory mechanism for arrest and initiation of primordial follicular growth is crucial for female fertility. Previously, we found 15 expressed sequence tags (ESTs) that were specifically abundant in the day-S-subtracted cDNA library and that the B357 clone was novel. The present study was conducted to obtain the whole sequence of the novel gene including B357 and to characterize its mRNA and protein expression in mouse ovary and testis. Methods: The extended sequence of the 2,965-bp cDNA fragment for the clone B357 was named ${\underline{5}}-{\underline{d}}ay-{\underline{o}}vary-{\underline{s}}pecific\;gene-{\underline{1}}$ (5DOS1) and submitted to GenBank (accession number ${\underline{AY751521}}$). Expression of 5DOS1 was characterized in both female and male gonads at various developmental stages by Northern blotting, real-time RT-PCR, in situ hybridization, Western blotting, and immunohistochemistry. Results: The 5DOS1 transcript was highly expressed in the adult testis, brain, and muscle as compared to the other tissues. In the ovary, the 5DOS1 transcript was detected in all oocytes from primordial to antral follicles, and highly expressed at day 5 after birth and decreased thereafter. In contrast, expression of 5DOS1 showed a gradual increase during testicular development and its expression was limited to various stages of male germ cells except spermatogonia. Conclusions: This is the first report on the expression and characterization of the 5DOS1 gene in the mouse gonads. Further functional analysis of the 5DOS1 protein will be required to predict its role in gametogenesis.

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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Mining Single Nucleotide Polymorphisms from Silkworm EST Data

  • Qingyou, Xia;Tingcai, Cheng;Jifeng, Qian;Zheyang, Zhou;Zhonghuai, Xiang
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.23-23
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    • 2003
  • We made use of 81, 635 expressed sequence tags (ESTs) derived from 12 different cDNA libraries of Bombyx mori to identify high-quality candidate single nucleotide polymorphisms (SNPs). By PHRAP assembling, we obtained 12, 980 contigs containing 11, 531 contigs assembled by more than one reads. From 117 contig sequences, which were assembled by 1, 576 high-quality reads base-called with PHRED, we identified 101 candidate SNPs and 27 single base insertions/deletions based on a neighborhood quality standard(NQS) of SNP. (omitted)

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The Brassica/Arabidopsis Comparative Genome Browser A Novel Approach to Genome Browsing

  • Lewis Christopher T.;Sharpe Andrew G.;Lydiate Derek J.;Parkin Isobel A.P.
    • Journal of Plant Biotechnology
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    • v.5 no.4
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    • pp.197-200
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    • 2003
  • Scalable Vector Graphics (SVG) has enabled a visually appealing, browser-based application for the display of Brassica sequences relative to Arabidopsis thaliana, and there are currently more than 70,000 B. napus Expressed Sequence Tags (ESTs) displayed. The client side of this browser is based on a Custom Graphical User Interface (CGUI) library which uses SVG, a new web graphics standard, to provide windowing functionality inside the web browser. This windowing functionality, combined with asynchronous data retrieval and client side rendering overcomes two of the key technology imposed drawbacks of current web based browsers: Fixed displays and frequent page reloads. The end result is an intuitive and enjoyable browsing experience. The browser is accessible online from the Brassica / Arabidopsis Genomics Initiative (http://brassica.agr.gc.ca). Inquiries about the browser should be directed to LewisCT@agr.gc.ca.

Molecular Cloning and Expression Patterning of Novel Gene in the Silk Gland from Larval Trichoptera

  • Eum, Jai-Hoon;Goo, Tae-Won;Yun, Eun-Young;Hwang, Jae-Sam;Kang, Seok-Woo;Han, Sung-Sik
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.52-52
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    • 2003
  • Expressed sequence tags(ESTs) constitute a rapid and informative strategy for studying gene-expression profiles of specific stages and tissues. In this report, a cDNA library constructed from late larval Hydropsyche sp. was used to generate ESTs. Caddis larval silk-gland produce silk which are used in constructing their retreats and cases in the aquatic situation. (omitted)

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Analysis of expressed sequence tags from Protaetia brevizarsis(Coleoptera).

  • Kang, Seok-Woo;Hong, Sun-Mee;Eum, Jai-Hoon;Goo, Tae-Won;Yun, Eun-Young;Park, Kwang-Ho;Hwang, Jae-Sam
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.66-66
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    • 2003
  • The coleoptera is the most species-rich order of animals. Relatively little is known about Coleoptera genes and genome. We describe here the construction and DNA sequencing of cDNA libraries from Protaetia brevizarsis, a fruit tree pest in Korea. We sequenced and analyzed 3072 ESTs from wholebody of Protaetia brevizarsis larvae. (omitted)

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Functional Analysis of ESTs from the Flower Bud 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.124-124
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    • 2003
  • In order to study gene expression in a reproductive organ, we constructed a cDNA library of immature flower buds in Korean ginseng and generated expressed sequence tags (ESTs) of 3,360 clones randomly selected. The ESTs could be clustered into 1,844 non-redundant groups. Similarity search of the non-redundant ESTs against public non-redundant databases of both protein and DNA indicated that 1,254 groups show similarity to genes of known function. These ESTs clones were divided into sixteen categories depending upon gene function. The most abundant transcripts were unknown protein (72), chlorophyll a/b-binding protein (48), and stylar glycoprotein. There are no useful informations of gene expression during the development of flower bud in Korean ginseng. These results could help to understand the development of flower bud in Korean ginseng.

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