• 제목/요약/키워드: ORF analysis

검색결과 293건 처리시간 0.028초

요각류 Paracyclopina nana Acetate Kinase의 클로닝 및 대장균에서의 발현 (Cloning of Acetate Kinase Gene from the Copepod Paracyclopina nana and its Expression in Escherichia coli)

  • 정상운;서정수;이영미;박태진;김일찬;박흠기;이재성
    • 미생물학회지
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    • 제41권3호
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    • pp.157-163
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    • 2005
  • 요각류 Paracyclopina nana Acetate Kinase를 클로닝하였다. 전체 open reading frame은 1,200 bp이었으며, poly(A) signal sequence가 ORF에 내재되어 있었다. 분자계통학적 분석결과 P. nana acetate kinase 유전자는 진핵생물계 곰팡이류인 Aspegillus와 같은 branch를 형성하였고, P. nana acetate kinase가 다른 원핵미생물들의 acetate kinase와는 구별되며 fungi와 같은 branch에 존재하는 것을 확인하였다. 또한, E. coli를 이용하여 원핵세포 발현벡터를 이용한 단백질 발현 유도를 통하여 P. nana acetate kinase 단백질 분자량이 약 50 kDa에 이르는 것을 확인하였다. 이 자료는 본 요각류와 다른 생물의 acetate kinase 단백질의 생화학적 특성비교에 유용하게 쓰이리라 사료된다.

여름느타리버섯으로부터 ${\beta}-tubulin$ cDNA의 분리 및 염기서열 결정 (Isolation and Sequencing of the cDNA Encoding ${\beta}-tubulin$ from Pleurotus sajor-caju)

  • 김범기;신평균;정미정;박수철;유영복;류진창;권석태
    • 한국균학회지
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    • 제25권1호통권80호
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    • pp.1-5
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    • 1997
  • 여름느타리버섯 균사체의 cDNA library로부터 ${\beta}-tubulin$ 유전자를 분리하여 염기서열을 분석하였다. 분리된 ${\beta}-tubulin$ cDNA유전자는 27nt의 5'-untranslation region과 1341nt의 open reading frame, 191nt의 3'-untranslation region으로 구성되어 있었다. ORF는 445개의 아미노산들로 구성되어 있으며, 동물, 식물, 사상균에서 보고된 ${\beta}-tubulin$과 80% 이상의 상동성을 보였다. 분리된 Myc7tub clone을 사용하여 Southern hybridization한 결과 여름느타리버섯에는 두 가지의 isotype ${\beta}-tubulin$이 존재할 것으로 생각된다.

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Molecular Characterization of Fusarium Graminearum Virus 2 Isolated from Fusarium graminearum Strain 98-8-60

  • Yu, Ji-Suk;Lee, Kyung-Mi;Son, Moon-Il;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • 제27권3호
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    • pp.285-290
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    • 2011
  • Fusarium graminearum virus 2 (FgV2) infects Fusarium graminearum strain 98-8-60 and has at least five segments of double-stranded RNAs (dsRNAs), denoted as dsRNA-1 to dsRNA-5. In this study, the genome of FgV2 was sequenced and its phylogenetic relationship with other mycoviruses was analyzed. The lengths of FgV2 dsRNAs 1-5 ranged from 2414 to 3580 base pairs (bp). The 5' and 3' untranslated regions (UTRs) are highly conserved, and each dsRNA segment had 78-105 and 84-306 bp of 5' and 3' UTRs, respectively. Each dsRNA segment contained a single open reading frame (ORF). Computer analysis of dsRNA-1 revealed a putative open reading frame (ORF) that shows high sequence identity with an RNA-dependent RNA polymerase (RdRp) containing eight conserved motifs. dsRNAs 2-5 also each contain one putative ORF coding for products of unknown function. The sequences of FgV2 dsRNA-2 and dsRNA-3 have significant sequence identity with Magnaporthe oryzae chrysovirus 1 (MoCV1) dsRNA-3 and -4, respectively. When compared to other dsRNA mycoviruses in a phylogenetic analysis of the putative RdRp protein, FgV2 was found to form a distinct virus clade with Aspergillus mycovirus 1816 and MoCV1 in the family Chrysoviridae.

LuxR-Type SCO6993 Negatively Regulates Antibiotic Production at the Transcriptional Stage by Binding to Promoters of Pathway-Specific Regulatory Genes in Streptomyces coelicolor

  • Tsevelkhoroloo, Maral;Li, Xiaoqiang;Jin, Xue-Mei;Shin, Jung-Ho;Lee, Chang-Ro;Kang, Yup;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • 제32권9호
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    • pp.1134-1145
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    • 2022
  • SCO6993 (606 amino acids) in Streptomyces coelicolor belongs to the large ATP-binding regulators of the LuxR family regulators having one DNA-binding motif. Our previous findings predicted that SCO6993 may suppress the production of pigmented antibiotics, actinorhodin, and undecylprodigiosin, in S. coelicolor, resulting in the characterization of its properties at the molecular level. SCO6993-disruptant, S. coelicolor ΔSCO6993 produced excess pigments in R2YE plates as early as the third day of culture and showed 9.0-fold and 1.8-fold increased production of actinorhodin and undecylprodigiosin in R2YE broth, respectively, compared with that by the wild strain and S. coelicolor ΔSCO6993/SCO6993+. Real-time polymerase chain reaction analysis showed that the transcription of actA and actII-ORF4 in the actinorhodin biosynthetic gene cluster and that of redD and redQ in the undecylprodigiosin biosynthetic gene cluster were significantly increased by SCO6993-disruptant. Electrophoretic mobility shift assay and DNase footprinting analysis confirmed that SCO6993 protein could bind only to the promoters of pathway-specific transcriptional activator genes, actII-ORF4 and redD, and a specific palindromic sequence is essential for SCO6993 binding. Moreover, SCO6993 bound to two palindromic sequences on its promoter region. These results indicate that SCO6993 suppresses the expression of other biosynthetic genes in the cluster by repressing the transcription of actII-ORF4 and redD and consequently negatively regulating antibiotic production.

High-level Expression, Polyclonal Antibody Preparation and Bioinformatics Analysis of Bombyx mori Nucleopolyhedrovirus orf47 Encodes Protein

  • Wu, Chao;Guo, Zhongjian;Chen, Keping;Shen, Hongxing
    • International Journal of Industrial Entomology and Biomaterials
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    • 제16권2호
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    • pp.87-92
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    • 2008
  • Bombyx mori nucleopolyhedrovirus (BmNPV) orf47 gene was characterized for the first time. The coding sequence of Bm47 was amplified and subcloned into the prokaryotic expression vector pET-30a(+) in order to produce His-tagged fusion protein in the BL21 (DE3) cells. The His-Bm47 fusion protein was expressed efficiently after induction with IPTG. The purified fusion protein was used to immunize New Zealand white rabbits to prepare polyclonal antibody. As the genome of BmNPV is available in GenBank and the EST database of BmNPV is expanding, identification of novel genes of BmNPV was conceivable by data-mining techniques and bioinformatics tools. Structural bioinformatics approach to analyze the properties of Bm47 encodes protein.

Molecular Cloning and Sequencing of Cell Wall Hydrolase Gene of an Alkalophilic Bacillus subtilis BL-29

  • Kim, Tae-Ho;Hong, Soon-Duck
    • Journal of Microbiology and Biotechnology
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    • 제7권4호
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    • pp.223-228
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    • 1997
  • A DNA fragment containing the gene for cell wall hydrolase of alkalophilic Bacillus subtilis BL-29 was cloned into E. coli JM109 using pUC18 as a vector. A recombinant plasmid, designated pCWL45B, was contained in the fragment originating from the alkalophilic B. subtilis BL-29 chromosomal DNA by Southern hybridization analysis. The nucleotide sequence of a 1.6-kb HindIII fragment containing a cell wall hydrolase-encoding gene was determined. The nucleotide sequence revealed an open reading frame (ORF) of 900 bp with a concensus ribosome-binding site located 6 nucleotide upstream from the ATG start codon. The primary amino acid sequence deduced from the nucleotide sequence revealed a putative protein of 299 amino acid residues with an M.W. of 33, 206. Based on comparison of the amino acid sequence of the ORF with amino acid sequences in the GenBank data, it showed significant homology to the sequence of cell wall amidase of the PBSX bacteriophage of B. subtilis.

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Development of Information Biology (III)

  • Tateno, Yoshio
    • Interdisciplinary Bio Central
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    • 제5권2호
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    • pp.5.1-5.3
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    • 2013
  • Introduced were two biological investigations in which information biology played a significant role. In the first case independent findings in cancer research over a long period were united and organized by information biology and led to the outcome that was subject to a Nobel Prize. The outcome has revealed that the cause of human cancer is located in the genome in a dormant condition. The second case shows how to elucidate the function of an unknown DNA sequence or ORF in prokaryotes by a large - scale computer homology search and analyses. For the elucidation the International DNA Databases and a large - scale computer were two key factors.

Genetic Engineering for Detection of Endocrine Disruption using I-18 C Gene Expression in Chironomus riparius

  • Kwak Inn-Sil
    • 환경생물
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    • 제23권3호
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    • pp.269-274
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    • 2005
  • The 2D/E gel analysis for polypeptide expression reflecting I-18 C gene (early-ecdysterone inducible gene) has conducted the emerged C. riparius adults from larval phase exposure to tebufenozide acting as an ecdysteroidal molting hormone. Control group, the amount of ORE II of the I-18 C gene was larger than that of ORE I of this gene. After treatments, ORE I of the I-18 C gene was overexpressed as the polypeptide, whereas ORF II of this gene was expressed as the polypeptide and was clearly reduced expression. Accordingly, we consider that tebufenozide exhibited endocrine disruptions related processing of ecdysteroid receptor protein reflecting ORF II of I-18 C gene. Also, earlier emergence day was related overexpressed polypeptide reflecting ORE I of I-18 C gene. In this study result, tebufenozide induced changing of physiological condition, and then polypeptide expression reflecting early-ecdysterone inducible I-18 C gene was different between control group and exposure group.

다중서열수집 및 변환을 위한 효과적인 바이오인포메틱스 도구 (An Effective Bioinformatics Tool for Multiple Sequence Acquisition and Translation)

  • 이혜리;이승희;이건명;김성수;이찬희;이성덕
    • 한국지능시스템학회논문지
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    • 제18권1호
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    • pp.27-31
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    • 2008
  • 많은 바이오인포매틱스 관련 데이터베이스와 도구가 네트워크를 통해서 제공되고 있고, 이들을 효과적으로 활용하면 생물학적 분석을 적은 비용으로 우수한 결과를 얻을 수 있다. 이 논문에서는 주어진 질의에 대해서 잠재적으로 관련된 DNA 서열 정보를 획득하고, 분석자가 관심 있는 항목을 선택하면, 선택된 항목에 대한 모든 DNA 서열 정보를 확보하고, 이들에 대해서 아미노산 서열로 자동변환하여 ORF라는 정보를 활용하여 가장 가능성이 큰 것을 추천하는 도구를 소개한다. 해당 도구에는 웹 로봇 기법과 ORF 검색등을 위한 생물학적 지식을 활용한다.

경북지역에서 분리된 Salmonella enterica Serovar Typhimurium의 Class I Integron Gene Cassette 특성과 PEGE 유형분석 (Structural Analysis of Class I Integron Gene Cassette and Assessment of Genetic Relationships by PFGE of Salmonella enterica Serovar Typhimurium Isolated in Gyeongbuk Area)

  • 손창규;이정아;이도영;허완;정중교
    • 미생물학회지
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    • 제42권1호
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    • pp.12-18
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    • 2006
  • 2003년부터 2004년까지 정북지역에서 발생한 설사환자로부터 17주, 돼지로부터 18주 등 총 35주의 Salmonella enterica serovar Typhimurium이 분리되었고, 분리된 균주를 대상으로 항균제 내성양상, class I integron 특성 및 pulse-field gel electrophoresis유형(PFGE 유형; pulsotype)이 조사되었다. 35주 모두가 한가지 이상의 항균제에 내성을 나타내었고, 돼지로부터 분리된 대부분의 균주는 ampicillin, chloramphenicol, streptomycin, sulfamethoxazole/trimethoprime, tetracyclin, nalidixic acid에 내성을 나타내었다. 35주를 대상으로 class I, II 및 III integron gene cassette를 검색한 결과, 설사환자로부터 분리된 17주 중 3주가 dhfrX-orfF-aadA2 integron gene cassette을 보유하였고, 돼지로부터 분리된 18주중 11주가 dhfrX-orfF-aadA2 integron gene cassette를, 1주가 aadA2 integron을 보유하였다. 그러나 35주 모두가 class 2 integron gene cassette와 class 3 integron gene cassette는 보유하지 않은 것으로 나타났다. 35주의 PEGE 유형은 5가지로 분류되었으며, 31주가 pulsotype A로 나머지 4주는 pulsotype B, C, D, E형으로 각각 나뉘어졌다. 이러한 결과로 볼 때 경북지역에서 사람과 돼지에서의 S. enterica serovar Typhimurium에 의한 살모넬라중은 몇 종류 유행주의 전파에 의한 것임을 보여줄다. dhfrX-orfF-aadA2 integron gene cassette를 보유한 13주가 pulsotype A, dhfrX-orfF-aadA2 integron gene cassette을 보유한 1주가 pulsotype B, aadA2 integron을 보유한 1주가 pulsotype E, integron을 보유하고 있지 않은 15주가 pulsotype A로 나타났다.