• Title/Summary/Keyword: Open Reading Frame

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Pseudosymptom Analysis Induced by Geminivirus L4 Gene in Transgenic Arabidopsis (Geminivirus L4 유전자 도입에 따른 형질전환 애기장대의 감염유사증상 분석)

  • Lee, Suk-Chan;Lee, Kyu-Bae;Park, Jong-Bum
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.129-133
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    • 2003
  • The factors for symptom development caused by Beat curly top virus(BCTV) have been analyzed by using a molecular genetic approach based on expressing BCTV encoded proteins in transgenic plants. BCTV open reading frame (ORF) L4 expression in transgenic Arabidopsis resulted in abnormal plant development and the production of callus inflorescence stems and bumpy trichomes, confiming that this gene alone is a primary symptom determinant. The L4 gene expression by northern hybridization in transgenic plants and a range of phenotypes were analyzed.

Insertional Mutation of the Rice Blast Resistance Gene, Pi-b, by Long Terminal Repeat of a Retrotransposon

  • Jwa, Nam-Soo;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.16 no.2
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    • pp.105-109
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    • 2000
  • The Pi-b is the rice gene conferring race specific resistance to the blast fungus Magnaporthe grisea race having a corresponding avirulence gene, AVR-Pi-b. All resistant cultivars have two copies of the Pi-b gene, but susceptible cultivars have a single copy of the gene. About 1 Kbp insertion sequence was detected in the open reading frame of the Pi-b gene from the susceptible cv. Nipponbare. The nature of insertion sequence was identified as a solo long terminal repeat (LTR) of new rice Tyl-copia-like retrotransposon. LTR was widely distributed in the rice genome. Various types of different patterns of restriction fragment length polymorphism of LTR were detected in indica cultivars, whereas a single type was detected from japonica cultivars. The insertion of LTR sequence in the Pi-b gene in the susceptible cultivar suggested that retrotransposon-mediated insertional mutation might played an important role in the resistance breakdown as well as evolution of resistance genes in rice.

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The 52 kD Protein Gene of Odontoglossum Ringspot Virus Containing RNA-Dependent RNA Polymerase Motifs and Comparisons with Other Tobamoviruses

  • Park, Won-Mok
    • Journal of Plant Biology
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    • v.38 no.2
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    • pp.129-136
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    • 1995
  • Complementary DNA of the genomic RNA of odontoglossum ringspot virus Cymbidium strain (ORSV-Cy) was synthesized from polyadenylated viral RNA and cloned. Selected clones containing the viral RNA-dependent RNA polymerase gene of the virus has been sequenced by automated sequencing system. The complete nucleotide sequence of an open reading frame is 1377 base pairs in length, and encodes a protein of 458 amino acids about 52, 334 D. The 52 kD protein of ORSV shares four sequence motifs characteristic of viral RNA-dependent RNA polymerase. Comparison of the ORSV 52 kD protein sequence with that of other five viruses in tobamovirus group showed 76.0 to 60.7% homologies at the amino acid level and the conservation of the four motifs betwen the viruses.

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Generation of polyclonal antiserum to olive flounder (Paralichthys olivaceus) immunoglobulin by immunization of rabbit with plasmids containing heavy chain gene of olive flounder immunoglobulin

  • Kim, Gi-Hong;Kwon, Se-Ryun;Kim, Chun-Soo;Lee, Eun-Hye
    • Journal of fish pathology
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    • v.19 no.2
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    • pp.183-188
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    • 2006
  • In fish vaccinology, the secondary antibodies against fish immunoglobulins (Igs) are necessary to measure specific humoral immune responses in immunized fish. In the present study, polyclonal antiserum against olive flounder (Paralichthys olivaceus) IgM heavy chain was generated by intramuscular immunization of rabbit with Escherichia coli/eukaryotic shuttle vector containing open reading frame (ORF) of olive flounder IgM heavy chain. Western blot analysis demonstrated the specific activity of the rabbit antiserum with reduced olive flounder serum H chain at dilutions up to 1:1000. Titer of immunized rabbit serum against olive flounder serum was significantly higher than that of pre-immunized rabbit serum when determined by ELISA.

Expression of Cytochrome $b_{5}$ Retropseudogenes in Hunam Blood

  • Hwang, Mi-Sun;Alan W.Steggles;Yoo, Min
    • Biomedical Science Letters
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    • v.9 no.3
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    • pp.167-170
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    • 2003
  • Cytochrome $b_{5}$($b_{5}$) can be found in a variety of tissues and plays a role in the electron transfer pathways. Several retropseudogenes (numbered as I, II, II, IV, V) have been identified and well investigated for their structures. However, retropseudogene I is not clear in terms of its location on the chromosome. In addition the structure and the exression of retropseudogene V have not been confirmed. To examine the structure of bs retropseudogenes V and to see whether it is expressed in human blood we applied recombinant DNA technologies including polymerase chain reaction (PCR) and DNA sequencing. Retropseudogene V turned out to contain open reading frame (ORF) within its structure, however, no evidence of its expression was detected. Retropseudogene I was also found on the chromosome V. This study should contribute to the understanding of the structure of bs gene family.

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Pseudomonas sp. strain DJ77 균주에서 extradiol dioxygenase 를 암호화하고 있는 phnE 유전자의 염기배열

  • 김영창;신명수;윤길상;박영순;김욱현
    • Korean Journal of Microbiology
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    • v.30 no.1
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    • pp.8-14
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    • 1992
  • Nucleotide Sequence of phnE Gene Encoding Extradiol Dioxygenase fromPseudomonas sp. Strain DJ77Kim, Young-Chang'.", Myeong-Su Shin1, Kil-Sang Younl, Young-Soon Park1, andUg-Hyeon Kim'.' (Department of Microbiology, C'hungbuk National University.Cheongju 360-763, KOREA. and 'Research Center for Molecular Microbiology,Seoul National University)nal University)

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Characterization of a gene encoding ornithine carbamoyltransferase from rice

  • Islam Sikdar, Shafiqul;Kim, Jung-Sup
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.397-402
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    • 2009
  • Ornithinine carbamoyltransferase (OTC) is an enzyme that catalyzes the key step in arginine biosynthesis in bacteria and plants. OTC is also involved in the urea cycle and deficiency of the enzyme in human leads to disease. The argF gene encoding OTC has been reported in many bacteria and few plants. Here we report the characterization of a gene encoding OTC from rice (OsOTC). Analysis of a cDNA sequence from rice revealed that the full-length open reading frame of OsOTC consisted of 367 amino acids, corresponding to a protein of approximately 39.7 kDa. The predicted amino acid sequence of OsOTC harbor distinct five OTC signature sites and is highly homologous to that of enzymes of plants, animals and many bacterial OTCs. Expression of OsOTC in argF mutants of Escherichia coli showed that the gene was able to functionally complement to the mutant. These results suggest that the OsOTC encode a protein for ornithine carbamoyltransferase in rice.

Molecular Cloning and Nucleotide Sequence of Amaranthus viridis Homologue of the H -Transporting ATPase Gene (비름에서 ATP 가수분해효소와 상동성을 가지는 유전자의 클로닝)

  • 한규웅
    • Journal of Life Science
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    • v.6 no.1
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    • pp.1-5
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    • 1996
  • Using differential hybridization, a cDNA clone was isolated fortuitously from Amaranthus viridis and sequenced. This nucleotide sequence exhibited 55.1% identity with vma6 which encodes the 36-kD subunit of the vacuolar proton transporting ATPase in Saccharmoyces cerevisiae. The predicted open reading frame encodes a protein of 221 amino acid sequence with a calculated molecular weight of 25,452 and reveals high levels of similarity with subunit D polypeptide of vacuolar H -ATP(e.g., 48.5, 52.1 and 49.3% identity to the vacuolar 36-kD chain of yeast, vacuolar 32-kD polypeptide IV of human and vacuolar 28-kD protein of bovine chromaffin granules, respectively). The hydropathy index computation revealed that this predicted protein is a peripheral protein. These results indicated that the predicted protein may play a sturctural role in the vaculor H -ATPase as does gamma subunit in V-type ATPase.

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Molecular Cloning of Red Seabream, Pagrus major Somatolactin cDNA and Its Expression in Escherichia coli

  • Munasinghe, Helani;Koh, Soon-Mi;Lee, Jehee
    • Journal of Aquaculture
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    • v.16 no.3
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    • pp.165-170
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    • 2003
  • Isolation, cloning and sequencing of red seabream (Pagrus major) somatolactin (rsbSL) cDNA from pituitary gland revealed an open reading frame of 693 bp coding for a pre-growth hormone of 231 amino acids with a 22 amino acid putative signal peptide. Deduced amino acid sequence showed that there was one possible N-glycosylation site at Asn$^{145}$ and seven Cys residues (Cys$_{29}$ , Cys$^{39}$ , Cys$^{66}$ , Cys$^{89}$ , Cys$^{205}$ , Cys$^{222}$ , Cys$^{230}$ ). Except Cys$^{66}$ , others may be involved in disulfide bond formation. The rsbSL presented a 93% amino acid sequence identity with the SL of gilthead seabream (Sparus aurata) and contained the conserved hormone domain region. Expression of rsbSL in E. coli (BL2l) cells and gel analysis revealed a higher molecular weight for rsbSL than expected theoretically, implying posttranslational modifications.

Isolation and Characterization of a CDNA Encoding a Protein Homologous to the Mouse 70 kDa Heat Shock Protein (생쥐 섬 유아세포에서 70 kDa 고온충격 단백질의 CDNA 클로닝과 염기서열 분석)

  • 김창환;정선미최준호
    • The Korean Journal of Zoology
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    • v.35 no.2
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    • pp.203-210
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    • 1992
  • Hsp70, a 70 kDa protein, is the maior protein expressed when cells are heat-shocked. A cDNA library from mouse ID13 cells was screened with the human hsp70 gene as a probe, and a positive clone was obtained. The positive clone was subcloned into puc19 and the precise restriction was obtained. The CDNA was sequenced by the Sanger's dideoxv termination method. Single open reading frame that codes for a protein of 70 kDa was found. The DNA sequence of the cloned mouse DNA shows great homology (66-90%) with other mouse hsp70 genes and somewhat less homology (50",) with E. coli hsp70 gene (dnak). With the exception of one amino acid, the protein sequence deduced from the CDNA is identical to the mouse that shock cognate protein 70 (hsc70) that is constitutivelv expressed at normal temperature. The result suggests that the cloned CDNA encodes a hsc70 family rather than a heatinducible family.mily.

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