• 제목/요약/키워드: gene conversion

검색결과 195건 처리시간 0.03초

Characterization of 2-hydroxymuconic semialdehyde dehydrogenase from Burkholderia cepacia G4

  • A. Matta Reddy;Min, Kyung-Rak;Kim, Young-Soo
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.218.2-219
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    • 2003
  • 2-Hydroxymuconic semialdehyde dehydrogenase catalyzes the conversion of 2-hydroxymuconic semialdehyde (HMS) to an enol form of 4-oxalocrotonate which is a step in the catechol-meta cleavage pathway. A tomC gene encoding 2-HMS dehydrogenase of Burkholderia cepacia G4, a soil bacterium that can grow on toluene, cresol, phenol or tricholoro ethylene, is identified in between catechol 2,3-dioxygenase gene and HMS hydrolase gene, its sequence is analysed and the enzyme is characterised. (omitted)

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Development of succinate producing Cellulomonas flavigena mutants with deleted succinate dehydrogenase gene

  • Lee, Heon-Hak;Jeon, Min-Ki;Yoon, Min-Ho
    • 농업과학연구
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    • 제44권1호
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    • pp.30-39
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    • 2017
  • This study was performed to produce succinic acid from biomass by developing mutants of Cellulomonas flavigena in which the succinate dehydrogenase gene (sdh) is deleted. For development of succinate producing mutants, the upstream and downstream regions of sdh gene from C. flavigena and antibiotic resistance gene (neo, bla) were inserted into pKC1139, and the recombinant plasmids were transformed into Escherichia coli ET12567/pUZ8002 which is a donor strain for conjugation. C. flavigena was conjugated with the transformed E. coli ET12567/pUZ8002 to induce the deletion of sdh in chromosome of this bacteria by double-crossover recombination. Two mutants (C. flavigena H-1 and H-2), in which sdh gene was deleted in the chromosome, were constructed and confirmed by PCR. To estimate the production of succinic acid by the two mutants when the culture broth was fermented with biomass such as CMC, xylan, locust gum, and rapeseed straw; the culture broth was analyzed by HPLC analysis. The succinic acid in the culture broth was not detected as a fermentation products of all biomass. One of the reasons for this may be the conversion of succinic acid to fumaric acid by sdh genes (Cfla_1014 - Cfla_1017 or Cfla_1916 - Cfla_1918) which remained in the chromosomal DNA of C. flavigena H-1 and H-2. The other reason could be the conversion of succinyl-CoA to other metabolites by enzymes related to the bypass pathway of TCA cycle.

Characterization of NAD-Dependent Formate Dehydrogenase from Trametes versicolor Using a Cell-Free Protein Expression System

  • LEE, Su-Yeon;JANG, Seokyoon;LEE, Soo-Min
    • Journal of the Korean Wood Science and Technology
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    • 제50권3호
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    • pp.159-166
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    • 2022
  • CO2 emissions are the primary reason for global warming; hence, biological and chemical technologies for converting CO2 into useful compounds are being actively studied. Biological methods using enzymes can convert CO2 under mild conditions. Formate dehydrogenase (FDH) is a representative CO2 conversion enzyme. Its function was revealed after isolation from bacteria, yeast, and plants. In this study, we evaluated the CO2 conversion potential of FDH isolated from wood-rotting fungi. After isolating the FDH gene (TvFDH) from Trametes versicolor, we cloned the full-length FDH from T. versicolor and expressed it in a cell-free expression system. The gene encoding TvFDH was identified as 1,200 bp open reading frame (ORF) and the expected molecular weight of the protein was approximately 42 kDa. Overexpression of the recombinant crude protein including TvFDH was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Enzyme activities and metabolite analyses confirmed the efficiency of TvFDH for CO2 reduction.

Discovery of Performance Traits-Linked Microsatellite Markers in Channel Catfish (Ictalurus punctatus)

  • Kim, Soon-Hag
    • 한국양식학회지
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    • 제18권2호
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    • pp.130-132
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    • 2005
  • Genomics research has two ultimate applied goals: to Isolate and clone genes of economic importance for bio-technology and gene-assisted selection (GAS), and to locate and use markers for marker-assisted selection (MAS) in selective breeding programs. To this end, we have identified linked markers for feed conversion efficiency growth rate, and disease resistance to enteric septicemia of catfish (ESC). Three microsatellite markers Ip266, Ip384, and Ip607 were identified to be linked to feed conversion efficiency. Similarly one marker each was identified to be linked to growth rate (Ip607) and disease resistance to ESC (Ip477). Ip607 marker linked to both growth rate and feed conversion efficiency, indicating that the QTL for both growth rate and feed conversion efficiency may either be the same or located in the same chromosomal region in the catfish genome. On phenotypic evaluation, certain traits such as growth rate can be accurately evaluated by body weight evaluation while other traits such as disease resistance can be quite complex. The linked DNA markers will be highly useful for MAS programs and for directing further efforts of genomic mapping for important quantitative traits.

담수환경에서 eDNA와 eRNA를 이용한 Microcystin 합성 남조류 탐색 및 세포 내 Microcystin 생합성 활성 변화 (Detection of Microcystin Synthetic Cyanobacteria and Variation of Intracellular Microcystin Synthesis Using by eDNA and eRNA in Freshwater Ecocystem)

  • 김건희;박채홍;조현진;권대률;황순진
    • 생태와환경
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    • 제56권1호
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    • pp.1-13
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    • 2023
  • 북한강 수역에서 가장 많이 검출되는 Microcystin (MC)을 대상으로 하여 MC 생합성 유전자(mcyA gene), 남조류 세포밀도, MC 농도 사이의 관계를 분석하여 RNA-MC 환산식을 도출하고 남조류 세포 내 존재하는 MC 농도를 예측하였다. 북한강 수역에서 mcyA 유전자는 묵현천 합류 이후 북한강 하류 지점에서 주로 발견되었으며 평균적으로 다른 지점보다 높은 copy number가 발견되었다. 북한강 상류 의암호 수역의 경우, 공지천 지점에서 mcyA 유전자 copy number가 증가하였으며 9월 이후 북한강 수역 전체에서 mcyA 유전자 copy number는 감소하였다. mcyA gene expression은 상류와 하류 수역의 시·공간적 차이가 존재하였으며 여름철 짧은 시기에 집중적으로 발현하였다. mcyA gene expression 양은 MC 농도와 상관성이 매우 높을 뿐만 아니라 MC을 생합성하는 것으로 알려진 Microcystis aeruginosa와 Dolichospermum circinale의 세포밀도와도 통계적으로 유의한 상관성이 존재하였다. RNA-MC 관계를 기반으로 도출된 6개의 환산식은 통계적 유의성을 보이며(p<0.05) 0.9 이상의 높은 상관계수(r)를 나타냈다. eRNA에 존재하는 MC 생합성 유전자 발현량은 수중의 남조독소 물질 합성을 판단하고 유전자의 활성 정도를 빠르게 정량하여 MC 발생 조기경보에 충분히 활용할 수 있을 것으로 판단된다.

Gene Expression and Secretion of the Anticoagulant Hirudin in Saccharomyces cerevisiae

  • Sohn, Jung-Hoon;Lee, Sang-Kwon;Choi, Eui-Sung;Rhee, Sang-Ki
    • Journal of Microbiology and Biotechnology
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    • 제1권4호
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    • pp.266-273
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    • 1991
  • Hirudin, a 65-amino acid protein isolated from the salivary gland of the bloodsucking leech, Hirudo medicinalis, is a potent thrombin-specific inhibitor and blocks the thrombin-mediated conversion of fibrinogen to fibrin in clot formation. We have studied the gene expression and secretion of hirudin in yeast. Saccharomyces cerevisiae. A gene coding for hirudin was synthesized based on the amino acid sequence and cloned into a yeast expression vector $YEG{\alpha}-1$ containing the ${\alpha}-mating$ factor pre-pro leader sequence and galactose-inducible promoter, GALl0. Recombinant S. cerevisiae was found to secrete biologically active hirudin into the extracellular medium. The secreted recombinant hirudin was recovered from the culture medium and purified with ultrafiltration and reverse phase high performance liquid chromatography. Approximately 1 mg of hirudin per liter was produced under suboptimal culture conditions and brought to about 90% purity in two steps of purification.

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Esherichia coli XMP Aminase 유전자의 발현 증대 (High Level Expression of XMP Aminase Gene in Esherichia coli)

  • 조정일;한철주
    • 한국식품위생안전성학회지
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    • 제6권3호
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    • pp.133-137
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    • 1991
  • 5'-XMP를 5'-GMP로 전환하는 효소인 XMP aminase[EC 6.3.4.1]의 활성을 증가시키기 위하여 XMP aminase의 유전자를 함유한 1.7kb gua A gene fragment를 pLC 34-10으로부터 분리하여 pBR 322에 subcloning 한 뒤 trp promoter를 가지고 있는 대장균 발현 벨터 pDR 720에 도입하였다. 재조합된 pXAR 64에 존재하는 gua A 유전자는 trp Promoter에 의하여 발현이 증대되었으며 $3-{\beta}-indoleacrylic$ acid에 의하여 XMP aminase의 생성이 유도되었다. XMP aminase의 비활성은 pLC 34-10을 함유한 균주에 비하여 약 17배 증가되었다.

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Isolation and Characterization of Kasugamycin Biosynthetic Genes from Streptomyces kasugaensis KACC 20262

  • JO YOU-YOUNG;LIU JING;JIN YING-YU;YANG YOUNG-YELL;SUH JOO-WON
    • Journal of Microbiology and Biotechnology
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    • 제15권3호
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    • pp.491-496
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    • 2005
  • The biosynthetic gene cluster for the aminoglycoside antibiotic kasugamycin was isolated and characterized from the kasugamycin producing strain, Streptomyces kasugaensis KACC 20262. By screening a fosmid library using kasA, the gene encoding aminotransferase, we isolated a 22 kb DNA fragment. The fragment contained seventeen complete open reading frames (ORFs); one of these ORFs, kasD, was identified as the gene for dNDP-glucose 4,6-dehydratase, which catalyzes the conversion of dNDP-glucose to 4-keto-6-deoxy-dNDP-glucose. The enzyme showed a broad spectrum of substrate specificity. In addition, ksR was overexpressed in E. coli BL21 and proved to be a self-resistance gene against kasugamycin. These findings suggest that the isolated gene cluster is highly likely responsible for the biosynthesis of kasugamycin.

인간 Y 염색체: 구조, 기능 그리고 진화 (Human Y Chromosome: Structure, Function and Evolution)

  • 홍경원;허재원;김희수
    • 생명과학회지
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    • 제13권6호
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    • pp.958-969
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    • 2003
  • 인간 Y 염색체는 엄격히 부계 유전되고 그 길이의 대부분은 남성 감수분열 동안 교차가 일어나지 않는다. 비록 이영역이 비 재조합 영역 Y (non-recombining region Y: NRY)로 불려왔지만, 풍부한 재조합의 발견으로 그것은 남성 특이 영역 (male-specific region Y: MSY)으로 재 명명(命名)되었다. MSY는 이질염색질 (heterochromatin) 서열과 세가지 분류의 진정염색질 (euchromatin) 서열 (X-전위영역, X-퇴화영역, 증폭영역)이 모자이크화 되어있다. X-전위영역의 서열은 X 염색체의 상동 좌위와 약 99% 동일성을 가진다. X-퇴화영역 서열은 고대 상 염색체가 현대의 X와 Y 염색체로 진화되면서 남아 있는 부분이다. 증폭영역의 8개의 회문구조는 인간 Y염색체의 남성 특이영역의 4분의 1을 차지한다. 이들은 많은 정소 특이 유전자를 포함하고, 회문구조서열 사이의 상동성은 약 99.97%이다. 이 회문구조의 양쪽 팔은 계속되는 유전자 교환에 의해 유지되며, 서로 협력하여 진화된다. 새로 태어나는 남성당 평균 약 600 염기당 하나가 Y-Y유전자 교환을 겪고, 정소 특이적 다중유전자군의 진화에 중요한 역할을 한다.

The Basis of Different Sensitivities of Ovarian Cancer Cells to the Recombinant Adenoviral Vector System Containing a Tumor-Specific L-plastin Promoter and E. coli Cytosine Deaminase Gene as a Transcription Unit

  • Chung, In-Jae
    • Biomolecules & Therapeutics
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    • 제17권2호
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    • pp.138-143
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    • 2009
  • We have reported previously on a replication incompetent recombinant adenoviral vector, AdLPCD, in which the expression of cytosine deaminase gene (CD) is driven by the tumor-specific L-plastin promoter. AdLPCD vector had been evaluated for its efficacy of chemosensitization of ovarian cancer cells to 5-FC. In spite of the fact that ovarian cancer cells, i.e., OVCAR-3 and SK-OV-3, are capable for adenoviral transduction judged by LacZ reporter gene analysis, two cell lines demonstrated quite different sensitivities toward AdLPCD/5-FC system. In OVCAR-3 cells, infection of AdLPCD followed by exposure to 5-FC resulted in the suppression of cell growth with statistical significance. On the other hand, SK-OV-3 cells were more resistant to the CD/5-FC strategy compared with OVCAR-3 cells under the same condition. The object of study was to investigate factors that would determine the sensitivity to AdLPCD/5-FC. We evaluated conversion rate of 5-FC to 5-FU after infection of AdLPCD by HPLC analysis, $IC_{50}$ of 5-FU, the expression level of integrin receptors i.e., ${\alpha}v{\beta}3$ and ${\alpha}v{\beta}5$, and status of p53 in OVCAR-3 and SK-OV-3 cells. The results indicated that OVCAR-3 cells have few favorable features compared with SK-OV-3 cells to be more effective to the AdLPCD/5-FC strategy; higher level of ${\alpha}v{\beta}5$ integrin, higher rate of conversion of 5-FC into 5-FC, and lower $IC_{50}$ of 5-FU. The results suggest that the replacement of 5-FU with CD/5-FC in combination chemotherapy would be less toxic and much greater cytotoxicity than the conventional combination chemotherapy in some patients.