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Characterization of RAD4 Homologous Gene from Coprinus cinereus (균류 Coprinus cinereus에서 DNA 회복에 관여하는 RAD4 유사유전자의 분리와 특성)

  • Choi, In-Soon
    • Journal of Life Science
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    • v.13 no.4
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    • pp.522-528
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    • 2003
  • The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the RAD4 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. In order to investigation whether the increase of transcripts by DNA damaging agent, transcripts levels were examined after treating the cells. The level of transcript did not increase by untraviolet light (UV). This result indicated that the RAD4 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the RAD4 homologous gene is essential for cell viability.

Isolation and Characterization of Cyclophilin 1 (ClCyP1) Gene from Codonopsis lanceolata (더덕의 주근에서 유래한 Cyclophilin 1 (ClCyP1) 유전자의 분리 및 분석)

  • 양덕춘;이강;인준교;이범수;김종학
    • Korean Journal of Plant Resources
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    • v.17 no.3
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    • pp.239-247
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    • 2004
  • A cyclophilin 1 cDNA clone(GenBank accession no.CF924191) was isolated from the taproot of C. lanceolata and designed as C1CyP1. Determination of the nucleotide sequence of C1CyPl identified an open reading frame of 525bp, which shared high homologies with cyclophilins that were previously reported in other organisms. The C1CyP1 amino acid sequence possesses 7 amino acid residue stretch(KSGKPLH) that is characteristic of plant cytosolic dehydrins. Currently available amino acid residues of plant cyclophilins were compared to examine their phylogenetic relationship to C1CyP1. In the phylogenetic analysis, based on the aligned sequences, C1CyP1 showed high homology with arabidopsis ROC2 and rice CyP1. The transcript that corresponded to C1CyP1 was abundant in callus, but only basal level of transcript was detected in stem, leaf and root. For the study in the defense mechanism against various stresses, we report expression patterns of this gene by quantative RT-PCR.

Drosophila CrebB is a Substrate of the Nonsense-Mediated mRNA Decay Pathway that Sustains Circadian Behaviors

  • Ri, Hwajung;Lee, Jongbin;Sonn, Jun Young;Yoo, Eunseok;Lim, Chunghun;Choe, Joonho
    • Molecules and Cells
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    • v.42 no.4
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    • pp.301-312
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    • 2019
  • Post-transcriptional regulation underlies the circadian control of gene expression and animal behaviors. However, the role of mRNA surveillance via the nonsense-mediated mRNA decay (NMD) pathway in circadian rhythms remains elusive. Here, we report that Drosophila NMD pathway acts in a subset of circadian pacemaker neurons to maintain robust 24 h rhythms of free-running locomotor activity. RNA interference-mediated depletion of key NMD factors in timeless-expressing clock cells decreased the amplitude of circadian locomotor behaviors. Transgenic manipulation of the NMD pathway in clock neurons expressing a neuropeptide PIGMENT-DISPERSING FACTOR (PDF) was sufficient to dampen or lengthen free-running locomotor rhythms. Confocal imaging of a transgenic NMD reporter revealed that arrhythmic Clock mutants exhibited stronger NMD activity in PDF-expressing neurons than wild-type. We further found that hypomorphic mutations in Suppressor with morphogenetic effect on genitalia 5 (Smg5) or Smg6 impaired circadian behaviors. These NMD mutants normally developed PDF-expressing clock neurons and displayed daily oscillations in the transcript levels of core clock genes. By contrast, the loss of Smg5 or Smg6 function affected the relative transcript levels of cAMP response element-binding protein B (CrebB) in an isoform-specific manner. Moreover, the overexpression of a transcriptional repressor form of CrebB rescued free-running locomotor rhythms in Smg5-depleted flies. These data demonstrate that CrebB is a rate-limiting substrate of the genetic NMD pathway important for the behavioral output of circadian clocks in Drosophila.

Expressional Evaluation of C/EBP Family, SREBP1, and Steroid Hormone Receptors in the Epididiymal Fat of Postnatally Developing Mouse

  • Lee, Yong-Seung;Lee, Ki-Ho
    • Development and Reproduction
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    • v.26 no.2
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    • pp.49-58
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    • 2022
  • The differentiation and development of preadipocyte into mature adipocyte are regulated by transcription factors, such as CCAAT enhancer binding protein (Cebp) gene family and sterol regulatory element binding transcription factor 1 (Srebp1). Steroid hormones give influences on the development and function of adipocyte. The present research examined expression patterns of CCAAT enhancer binding protein alpha (Cebpa), CCAAT enhancer binding protein beta (Cebpb), CCAAT enhancer binding protein gamma (Cebpg), sterol regulatory element binding transcription factor 1 (Srebp1), androgen receptor (Ar), and estrogen receptors (Esr) among different epididymal fat parts during postnatal period by quantitative real-time polymerase chain reaction. In the distal epididymal fat, expression of Cebpa, Cebpb, Cebpg, Srebp1, Ar, and Esr2 was increased until 12 months of age, while expression of Esr1 was decreased at 5 months of age and was not detectable after 8 months of age. In the proximal epididymal fat, transcript levels of Cebps and Srebp1 were increased at 8 months of age, followed by decreases of Cebpb and Cebpg transcript levels at 12 months of age. An additional increase of Srebp1 expression was observed at 12 months of age. Expression of Ar and Esr2 were increased until 8 months of age, followed by a drop of Ar expression level at 12 months of age. Expression pattern of Esr1 was similar to that in the distal epididymal fat. In the tail epididymal fat, expression of Cebpa, Cebpg, Srebp1, Ar, and Esr2 was increased with age. Esr1 was not detectable at all. The highest level of Cebpb was observed at 8 months of age. These data suggest the possibility of developmental and functional differentiation among the epididymal fat parts.

Genomic Analysis of Satellite RNA of Cucumber mosaic virus-Paf Related with Mild Symptoms (Cucumber mosaic virus Paf 계통의 약독 병징과 관련된 satellite RNA의 유전자 해석)

  • Sung, Mi-Young;Jung, Min-Young;Lee, Sang-Yong;Ryu, Ki-Hyun;Choi, Jang-Kyung
    • Research in Plant Disease
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    • v.10 no.4
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    • pp.241-247
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    • 2004
  • Recently, we reported a satellite RNA (Paf-satRNA) which is encapsidated in a pepper isolate of Cucumber mosaic virus (CMV-Paf) regulated symptom attenuation of the helper virus. To characterize mild symptom domain of Paf-satRNA, a series of chimeric cDNAs of satRNAs were created by using full-length cDNA clones of Paf-satRNA and a Pep-satRNA, chlorosis-inducing satRNA in pepper plants, and analyzed for determinants of symptom attenuation. When compared the nucleotide sequences, the 3' and 5' terminal sequences of the two wild-type (wt) satRNAs contained relatively conserved sequences which are the typical to CMV satRNA. Ten bases insertions were found in PepY-satRNA, and two variable regions, 81st to 113th and 183rd to 265th from the 5'-end, were located in the middle parts of the satRNAs. To delineate the attenuated symptom-related domain for the Paf-satRNA, in vitro transcripts RNAs transcribed from the wt cDNAs and constructed chimeric cDNAs were combined with genomic RNAs, RNA1, RNA2 and RNA3, of CMV-Fny and inoculated onto Nicotiana benthamiana plants. These transcripts were fully infectious onto the N. benthamiana and infectivity was confirmed by the RT-PCR. Chimeric Paf(H/N)-satRNA and PepY(N/A)-satRNA as well as Paf-satRNA induced very mild mosaic or symptomless infection on N. benthamiana. By contrast, typical mosaic symptom and stunting of infected plants were induced when PepY-satRNA, PepY(H/N)-satRNA and Paf(N/A)-satRNA were infected to N. benthamiana. Paf-satRNA coinfected with CMV-Fny RNAs induced very mild to sympomless on pepper plants whereas PepY-satRNA-infected pepper expressed typical chlorosis mosaic symptom. Two kinds of chimeric mutants, Paf(H/N)-satRNA and PepY(N/A)-satRNA, induced mild mosaic or symptomless infection onto pepper plants, while PepY(H/N)-satRNA and Paf(N/A)-satRNA showed typical chlorosis and mosaic symptom with stunting. This results suggest that mild symptom-related domain for the Paf-satRNA was located on HpaI-NarI region.

Spliced leader sequences detected in EST data of the dinoflagellates Cochlodinium polykrikoides and Prorocentrum minimum

  • Guo, Ruoyu;Ki, Jang-Seu
    • ALGAE
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    • v.26 no.3
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    • pp.229-235
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    • 2011
  • Spliced leader (SL) trans-splicing is a mRNA processing mechanism in dinoflagellate nuclear genes. Although studies have identified a short, conserved dinoflagellate SL (dinoSL) sequence (22-nt) in their nuclear-encoded transcripts, whether the majority of nuclear-coded transcripts in dinoflagellates have the dinoSL sequence remains doubtful. In this study, we investigated dinoSL-containing gene transcripts using 454 pyrosequencing data (Cochlodinium polykrikoides, 93 K sequence reads, 31 Mb; Prorocentrum minimum, 773 K sequence reads, 291 Mb). After making comparisons and performing local BLAST searches, we identified dinoSL for one C. polykrikoides gene transcript and eight P. minimum gene transcripts. This showed transcripts containing the dinoSL sequence were markedly fewer in number than the total expressed sequence tag (EST) transcripts. In addition, we found no direct evidence to prove that most dinoflagellate nuclear-coded transcripts have this dinoSL sequence.

An Agent Based IP Transcript System in VoIP Network (VoIP망에서 Agent 기반 IP 녹취 시스템)

  • Lim Jae-Jin;Kim Soo-Hee;Jung In-Sang;Jung In-Hwan
    • Proceedings of the Korea Information Processing Society Conference
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    • 2006.05a
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    • pp.1243-1246
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    • 2006
  • 초고속 통신망의 확대 적용으로 인터넷의 빠른 성장과 함께 음성과 비디오 그리고 데이터를 통합하고자 하는 노력이 시도되고 있다. VoIP(Voice over IP)는 IP를 이용하여 음성과 데이터를 패킷 형태로 통합하여 실시간으로 전송하는 기술이다[1]. 패킷 네트워크에서 VoIP 시그널링 기술을 이용하면 망 자원으 효율적 이용 및 PSTN에 가까운 음질 그리고 인터넷과 연계한 다양한 음성서비스 지원이 가능하다. 콜센터에서도 VoIP를 사용하게 됨에 따라 VoIP망에서의 녹취 시스템이 필요하다. VoIP 녹취 시스템은 상담원과 고객 간의 통화 내용을 자동으로 녹음하여 보관함으로써 고객의 요구사항을 명확하게 파악할 수 있으며 녹취 데이터의 통계 자료 제공으로 효율적인 관리가 가능하고, 선택 녹취, 스케쥴링 녹취, 상담원의 평가 자료를 제공하여 고객 관리의 질적인 향상을 지원한다. 본 논문에서는 성능에 큰 영향을 주지 않고 기존의 VoIP 녹취 시스템의 문제점을 해결한 에이전트를 포함한 VoIP 녹취 시스템을 제안한다.

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An Linux Based IP Transcript System in VoIP Network (VoIP망에서 리눅스기반 IP 녹취 시스템 설계)

  • Kim Soo-Hee;Kim Jin-Hwan;Jung In-Hwan
    • Proceedings of the Korea Information Processing Society Conference
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    • 2006.05a
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    • pp.1235-1238
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    • 2006
  • VoIP(Voice over IP)는 IP를 이용하여 음성과 데이터를 패킷 형태로 통합하여 실시간으로 전송하는 기술이다[1]. 본 논문에서는 VoIP망에서 리눅스 기반 IP 녹취 시스템을 설계 및 구현한다. 녹취 시스템은 고객과 상담원의 통화 내용을 자동으로 녹음하여 보관함으로써 고객의 요구 사항을 명확히 파악할 수 있다. 녹취 시스템으로 모든 네트워크 환경에서 사용할 수 있으며 CTI와 연동하여 효율적이고 체계적인 녹취 시스템 구축이 가능하다.

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Identification of Differentially Expressed Genes by cDNA-AFLP in Magnaporthe oryzae

  • Chi, Myoung-Hwan;Park, Sook-Young
    • Research in Plant Disease
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    • v.25 no.4
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    • pp.205-212
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    • 2019
  • Analysis of differentially expressed genes has assisted discovery of gene sets involved in particular biological processes. The purpose of this study was to identify genes involved in appressorium formation in the rice blast fungus Magnaporthe oryzae via analysis of cDNA-amplified fragment length polymorphisms. Amplification of appressorial and vegetative mycelial cDNAs using 28 primer combinations generated over 200 differentially expressed transcript-derived fragments (TDFs). TDFs were excised from gels, re-amplified by PCR, cloned, and sequenced. Forty-four of 52 clones analyzed corresponded to 42 genes. Quantitative real-time PCR showed that expression of 23 genes was up-regulated during appressorium formation, one of which was the MCK1 gene that had been shown to be involved in appressorium formation. This study will be providing valuable resources for identifying the genes such as pathogenicity-related genes in M. oryzae.

Cloning of Epidermis-specific cDNAS Encoding a Lipid Transfer Protein and an Aldehyde Decarbonylase from Senecio odorus

  • Pyee, Jaeho
    • Journal of Plant Biology
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    • v.39 no.3
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    • pp.189-195
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    • 1996
  • The major cuticular components have been shown to be synthesized in the epidermis. Therefore, cloning of epidermis-specific genes could yield information to be used to isolate and characterize the enzymes involved in the cuticle biosynthesis. A subtractive cDNA library was prepared from Senecio odorus in which epidermis-specific cDNAs were enriched. Differential screening of the library using epidermal and non-epidermal probes revealed two cDNAs. One of them designated epi425 was identified, based on the sequence homology, as a member of a new class in the LTP gene family and the other clone designated epi23 as a gene encoding an aldehyde decarbonylase. Northern blot analyses showed that epi425 and epi23 cDNAs hybridized with a transcript of about 600 and 2, 100 nucleotides, respectively, from the epidermis but not from the non-epidermal tissues. Further characterization of these clones will provide more information on the mechanism of the cuticle biosynthesis.

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