• Title/Summary/Keyword: Operator DNA

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A Vision Based Bio-Cell Recognition for Biomanipulation with Multiple Views

  • Jang, Min-Soo;Lee, Seok-Joo;Lee, Ho-Dong;Kim, Byung-Kyu;Park, Jong-Oh;Park, Gwi-Tae
    • 제어로봇시스템학회:학술대회논문집
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    • 2003.10a
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    • pp.2435-2440
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    • 2003
  • Manipulation of the nano/micro scale object has been a key technology in biology as the sizes of DNA, chromosome, nucleus, cell and embryo are within such order. For instance, for embryo cell manipulation, the cell injection is performed manually. The operator often spends over a year to carry out a cell manipulation project. Since the typical success rate of such operation is extremely low, automation of such biological cell manipulation has been asked. As the operator spends most of his time in finding the position of cell in the Petri dish and in injecting bio-material to the cell from the best orientation. In this paper, we propose a new strategy and a vision system, by which one can find, recognize and track nucleus, polar body, and zona pellucida of the embryo cell for automatic biomanipulation. The deformable template matching algorithm has been used in recognizing the nucleus and polar body of each cell. Result suggests that it outperforms the conventional methods.

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Modification of Animal Genotypes for the Regulation of Transgene Expression (이식유전자 발현조절을 위한 동물유전자의 조작)

  • 진동일
    • Korean Journal of Animal Reproduction
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    • v.19 no.4
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    • pp.283-291
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    • 1996
  • Transgenic animal을 응용할 수 있는 분야에서는 이식유전자의 기능을 정확하게 규명하고 이를 바탕으로 실질적인 유전적인 개량을 이루기 위해서 이식유전자의 발현을 조절할 수 있는 정교한 system이 필요하다. 유전자의 미세주입법에 의해 transgenic animal을 생산할 수 있는데 이용되고 있는 tissue-specific promoter에 의한 이식유전자의 발현조절은 필요로 하는 시기나 양 등을 인위적으로 조절하고자 하는데 한계점을 갖고 있다. 이러한 이식유전자 발현의 문제점을 극복하기 위해 효모의 recombinase나 미생물의 repressor 단백질과 이들의 binding site인 operator sequence를 이용하여 인위적으로 이식유전자의 발현을 조절할 수 있는 system이 개발되고 있다. Cre/loxP system은 site-specific recombination에 의해 DNA sequence를 제거함으로서 이식유전자의 발현을 조절할 수 있다. 이식유전자 발현의 장소와 양을 조절하기 위해서는 미생물이 이용하고 있는 repressor와 이들의 operator sequence를 적용하여 ligand binary system이 개발되었다. Lac repressor system에서는 isopropyl-$\beta$-D-thiogalactoside (IPTG)가 이식유전자 발현을 조절할 수 있는 positive regulator로서 작용하고, tetracycline-VP16 system에서는 tetracycline이나 유사물질들이 negative regulator로서 이용할 수 있다. 이러한 binary system은 transgenic animal에서 이식유전자 발현의 장소와 시기 또한 양을 효과적으로 조절하는데 적용할 수 있는 것으로 나타났다. 따라서 기존의 binary system과 함께 새로운 regulatory system의 장점을 이용하여 보다 완벽한 이식유전자의 인위적인 조절 system을 이룩함으로서 transgenic animal technology의 실용화를 앞당길 것으로 기대된다.

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Moderately thermostable phage Φ11 Cro repressor has novel DNA-binding capacity and physicochemical properties

  • Das, Malabika;Ganguly, Tridib;Bandhu, Amitava;Mondal, Rajkrishna;Chanda, Palas K.;Jana, Biswanath;Sau, Subrata
    • BMB Reports
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    • v.42 no.3
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    • pp.160-165
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    • 2009
  • The temperate Staphylococcus aureus phage ${\Phi}11$ harbors cI and cro repressor genes similar to those of lambdoid phages. Using extremely pure ${\Phi}11$ Cro (the product of the ${\Phi}11$ cro gene) we demonstrated that this protein possesses a single domain structure, forms dimers in solution at micromolar concentrations and maintains a largely $\alpha$-helical structure even at $45^{\circ}C$. ${\Phi}11$ Cro was sensitive to thermolysin at temperatures ranging from $55-75^{\circ}C$ and began to aggregate at ${\sim}63^{\circ}C$, suggesting that the protein is moderately thermostable. Of the three homologous 15-bp operators (O1, O2, and O3) in the ${\Phi}11$ cI-cro intergenic region, ${\Phi}11$ Cro only binds efficiently to O3, which is located upstream of the cI gene. Our comparative analyses indicate that the DNA binding capacity, secondary structure and dimerization efficiency of thermostable ${\Phi}11$ Cro are distinct from those of P22 Cro and $\lambda$ Cro, the best characterized representatives of the two structurally different Cro families.

The Binding Affinities of Two Binding Sites(CRP1 and CRP2 Sites) by Cyclic AMP Receptor Protein at Lactose Operon (락토스 오페론에서 Cyclic AMP Receptor Protein에 의한 두 결합 부위(CRP1과 CRP2)의 결합 특성에 관한 연구)

  • Kang, Jong-Baek;Kwon, Gun
    • Journal of Life Science
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    • v.13 no.5
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    • pp.746-750
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    • 2003
  • Lactose operon contains two CRP binding sites at promoter(CRP1 site) and operator(CRP2 site) regions at lac operon. CRP protein can bind to both sites with the different binding affinity. CRP1 site, major CRP binding site, acts the transcription activation with the fully unknown mechanism by binding of CRP. In this study, the binding affinities of CRP1 site and CRP2 site were measured with the fluorescein-labeled oligomers, which contain CRP1 site and the three different spacing sequences between GTGA and TCAC at CRP2 site. Results showed that CRP:cAMP complex bound to CRP1 site 3 times more strongly than CRP2 site and the base spacing between GTGA and TCAC was not the only factor to affect the binding affinity of CRP to CRP2 site.