• Title/Summary/Keyword: Cell Transformation

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Production of ρ-Hydroxyacetophenone by Engineered Escherichia coli Heterologously Expressing 1-(4-Hydroxyphenyl)-Ethanol Dehydrogenase

  • Wenmei Wu;Xiwei Yuan;Xin Gao;Chaoyang Tan;Shunxiang Li;Dehong Xu
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.467-475
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    • 2024
  • ρ-Hydroxyacetophenone is an important and versatile compound that has been widely used in medicine, cosmetics, new materials, and other fields. At present, there are two ways to obtain ρ-hydroxyacetophenone. One is to extract it from plants, such as Artemisia capillaris Thunb and Cynanchum otophyllum Schneid, and the other is to synthesize it by using chemical methods. Of these two methods, the second is the main one, although it has problems, such as flammable and explosive reagents, difficult separation of by-products, and harsh reaction conditions. To solve these issues, we adopted genetic engineering in this study to construct engineered Escherichia coli containing Hped gene or EbA309 gene. Whole-cell biotransformation was conducted under the same conditions to select the engineered E. coli with the higher activity. Orthogonal tests were conducted to determine the optimal biotransformation condition of the engineered E. coli. The results showed that the optimal condition was as follows: substrate concentration of 40 mmol/l, IPTG concentration of 0.1 mmol/l, an induction temperature of 25℃, and a transformation temperature of 35℃. Under this condition, the effects of transformation time on the ρ-hydroxyacetophenone concentration and cell growth were further studied. We found that as the transformation time extended, the ρ-hydroxyacetophenone concentration showed a gradually increasing trend. However, when the ρ-hydroxyacetophenone concentration increased to 1583.19 ± 44.34 mg/l in 24 h, cell growth was inhibited and then entered a plateau. In this research, we realized the synthesis of ρ-hydroxyacetophenone by biotransformation, and our findings lay a preliminary foundation for further improving and developing this method.

Introduction of Hog Cholera Virus Gene into Potato Plants by Agrobacterium-mediated Transformation and the Analysis of Its Expression

  • Kim, Hyun-Soon;Jeon, Jae-Heung;Kim, Cheol-Jung;Hyouk Joung
    • Journal of Plant Biotechnology
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    • v.4 no.4
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    • pp.155-161
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    • 2002
  • The HCV gene was expressed in potato plants under the control of the constitutive CaMV 355 promoter or tuber-specific patatin promoter. Solanum tuberosum plants carrying a plant expression vector harboring the encoding region of HCV gene were generated by Agrobacterium tumefaciens-mediated in vitro transformation methods. The presence of HCV gene in the plant genome was detected by PCR and DNA hybridization experiments. We obtained the 5 lines of transgenic potato with the pMBPHCV construct and 4 lines of transgenic potato with the pATHCV construct. The HCV transgenic stably integrated into the potato genome, as well as their transcription. HCV mRNA was identified in leaf and tuber tissues of transgenic plants by Northern blot analysis. The transgenic potato plants produced the expected transcript, and the corresponding HCV protein accumulated in individual transgenic plants.

Malignant Transformation of an Epidermoid Cyst in the Cerebellopontine Angle

  • Chon, Kyu-Hyon;Lee, Jong-Myong;Koh, Eun-Jung;Choi, Ha-Young
    • Journal of Korean Neurosurgical Society
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    • v.52 no.2
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    • pp.148-151
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    • 2012
  • Intracranial squamous cell carcinoma is extremely rare, with most of the cases arising from malignant transformation of an epidermoid or a dermoid cyst. The patient presented with facial weakness. Initial magnetic resonance imaging revealed a mass in the right cerebellopontine angle. A subtotal resection was performed via right retrosigmoid suboccipital approach. Histopathological findings were consistent with an epidermoid tumor. Five months later, the patient underwent gamma knife radiosurgery due to highly probable recurrent epidermoid tumor. Two years after, the patient's neurological deficit had been newly developed, and follow-up magnetic resonance imaging demonstrated a large contrast-enhancing tumor in the left cerebellopontine angle, which compressed the brainstem. After resection of the tumor, histopathological examinations revealed a squamous cell carcinoma probably arising from an underlying epidermoid cyst. We report a case of an epidermoid tumor in the cerebellopontine angle that transformed into a squamous cell carcinoma.

Optimal Volume Estimation for Non-point Source Control Retention Considering Spatio-Temporal Variation of Land Surface (지표면의 시공간적 변화를 고려한 비점오염원 저감 저류지 최적용량산정)

  • Choi, Daegyu;Kim, Jin Kwan;Lee, Jae Kwan;Kim, Sangdan
    • Journal of Korean Society on Water Environment
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    • v.27 no.1
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    • pp.9-18
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    • 2011
  • In this study the optimal volume for non-point source control retention is estimated considering spatio-temporal variation of land surface. The 3-parameter mixed exponential probability density function is used to represent the statistical properties of rainfall events, and NRCS-CN method is applied as rainfall-runoff transformation. The catchment drainage area is divided into individual $30m{\times}30m$ cells, and runoff curve number is estimated at each cell. Using the derived probability density function theory, the stormwater probability density function at each cell is derived from the rainfall probability density function and NRCS-CN rainfall-runoff transformation. Considering the antecedent soil moisture condition at each cell and the spatial variation of CN value at the whole catchment drainage area, the ensemble stormwater capture curve is established to estimate the optimal volume for an non-point source control retention. The comparison between spatio-temporally varied land surface and constant land surface is presented as a case study for a urban drainage area.

A Phase Transformation Study on Amorphous Diopside ($CaMgSi_2O_6$) (비정질 투휘석($CaMgSi_2O_6$)에 대한 상변이 연구)

  • 김영호
    • Journal of the Mineralogical Society of Korea
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    • v.16 no.2
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    • pp.161-169
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    • 2003
  • A phase transformation study on a synthetic amorphous diopside, $(Ca,Mg)SiO_3$has been carried out up to ∼30 GPa, and ∼$1000^{\circ}C$ using a diamond anvil cell and YAG laser heating system, respectively. A starting amorphous material shows a direct transition to cubic $(Ca,Mg)SiO_3$perovskite at high pressure, which contradicts to the crystalline diopside phase transformation sequence disproportionating into mixtures of the orthorhombic$ MgSiO_3$perovskite and the cubic $CaSiO_3$perovskite phases. This discrepancy might be due to the different starting materials as well as the temperature variations at each specific experiment performed. The present phase transfor mation sequence would modify the mineralogical assemblage in the Earth transition region and the lower mantle depending upon the pressure, temperature and the oxygen partial pressure.

Introduction of Ti Plasmid into Bradyrhizobium japonicum by Spheroplast Transformation (형질전환(形質轉換)에 의한 Ti Plasmid의 Bradyrhizobium japonicum에의 도입(導入))

  • Yun, Han-Dae;Cho, Moo-Je;Park, Chan-Young;Lee, Ke-Ho
    • Applied Biological Chemistry
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    • v.30 no.4
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    • pp.293-299
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    • 1987
  • Bradyrhizobium japonicum spheroplasts were prepared by culturing cells in the presence of glycine, follwed by treatment with lysozyme. The cells were examined by electron microscopy during the formation of spheroplast. Then Ti plasmid from Agrobacterium tumefaciens 15955 was introduced into Bradyrhizobium japonicum by glycine-lysozyme induced spheroplast transformation. After cell wall regeneration, transformants were selected by the ability of utilization of octopine. Transformation were received at a frequency of $1{\times}10^{-7}$. The transformants obtained from spheroplast transformation harbored the introduced Ti plasmid, which was identified by agarose gel electrophoresis. Furthermore, the differences in their gene products were observed between the transformant and the recipient cell by two-dimensional polyacrylamide gel electrophoresis. The transformants which still possessed the same ability nodulate soybean (Glycine max.) as that of the original host strain, acquired the ability to induce tumor on Petunia hybrida like Agrobacterium, but formed the small crown galls in size compared to those of Agrobacterium tumefaciens.

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The Use of Glufosinate as a Selective Marker for the Transformation of Cucumber (Cucumis sativus L.) (오이의 형질전환을 위반 선발마커로서 Glufosinate의 이용)

  • Cho Mi-Ae;Song Yun-Mi;Park Yun-Ok;Ko Suck-Min;Min Sung-Ran;Liu Jang-Ryol;Choi Pil-Son
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.161-165
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    • 2005
  • Agrobacterium tumefaciens-mediated cotyledonary explants transformation was used to produce transgenic cucumber. Cotyledonary explants of cucumber (c.v., Eunchim) were co-cultivated with strains Agrobaderium (LBA4404, GV3101, EHA101) containing the binary vector (pPTN289) carrying with CaMV 355 promoter-gus gene as reporter and NOS promoter-bar gene conferring resistance to glufosinate (herbicide Basta) as selectable marker. There was a significant difference in the transformation frequency depending Agrobacterium strains. The EHA101 of bacterial strains employed gave the maximum frequency (0.35%) for cucumber transformation. Histochemical gus and leaf painting assay showed that 15 individual lines were transgenic with the gus and bar gene. Southern blot analysis also revealed that the gus gene was successfully integrated into each genome of transgenic cucumber.

The Effect of Honey Bee Venom Aqua-Acupuncture by its Production from Different Regions on Decreased Immune Response (산지별(産地別) 봉독액약침자극(峰毒液藥鍼刺戟)이 면역기능저하(免疫機能低下)에 미치는 영향(影響))

  • Lee, Kyung-Hee;Kim, Chang-Hwan;Kang, Sung-Keel;Koh, Hyung-Kyun
    • Journal of Acupuncture Research
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    • v.17 no.4
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    • pp.28-40
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    • 2000
  • Objectives : This study was purposed to investigate the effect of honey bee venom aqua-acupuncture on decreased immune response by its production from Korea, North America and China. Methods : ICR-mouse was exposed with four minute sublethal 60Co- $\gamma$ -ray irradiation onto the body and got a series of each country's 0.1 ml honey bee venom aqua-acupuncture to Choksamni(ST36) every other day for three times. Thereafter the numbers of WBC, delayed type hypersensitivity, hemagglutinin titers, hemolysin titers, quantitation of T-cell and B-cell, lymphocyte transformation, natural killer cell activity, interleukin-2 productivity and interferon productivity were measured. Results : The numbers of WBC, delayed type hypersensitivity, hemagglutinin titers, hemolysin titers, quantitation of T-cell and B-cell and lymphocyte transformation were increased respectively with statistical significance in Korean, North American and Chinese honey bee venom aqua-acupuncture groups as compared with the control group. The natural killer cell activity was increased with statistical significance in Korean honey bee venom aqua-acupuncture group, and Interleukin-2 productivity and interferon productivity were not shown any statistical significance in all experimental groups as compared with the control group. Conclusions : According to the results, honey bee venom aqua-acupuncture has significant effect on decreased immune response, and Korean bee venom can be substituted for North American and Chinese one.

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Production of hGM-CSF from Cell Suspension Culture of Transformed Lettuce Using Agrobacterium-mediated Transformation System (Agrobacterium을 이용한 형질전환 상추의 세포 현탁배양으로부터 hGM-CSF의 생산)

  • Kim, Young-Sook;Kim, Mi-Young;Kwon, Tae-Ho;Yang, Moon-Sik
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.97-102
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    • 2003
  • Lettuce (Lactuca sativa) was transformed with Agrobacterium tumefacience LBA4404 containing human granulocyte macrophage colony stimulating factor (hGM-CSF) gene to produce in cell suspension cultures. Cell suspension culture was established using callus from transgenic lettuce plant. Integration of hGM-CSF gene into plant chromosome was confirmed through genomic PCR and Southern blot analysis. In addition, Northern blot analysis indicated the expression of the introduced hGM-CSF gene in transformed lettuce. The recombinant hGM-CSF was expressed in transgenic cell cultures derived from transgenic plants as a yield of about 149.0 $\mu\textrm{g}$/L in culture filtrate, which was determined by ELISA. These results demonstrated that transformed lettuce cell suspension cultures could be used as a production system of therapeutic proteins such as hGM-CSF.

Phase Transformation of Poly(trimethylene terephthalate) in Crystalline State: An Atomistic Modeling Approach

  • Jang, Seung-Soon;Jo, Won-Ho
    • Fibers and Polymers
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    • v.1 no.1
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    • pp.18-24
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    • 2000
  • The phase transformation of poly(trimethylene terephthalate) in crystalline state was simulated by atomistic modeling using molecular mechanics technique. The crystalline structure of PTT was successfully prepared using the well-defined unit cell structure of PTT and was satisfactorily verified by comparing that with the structure obtained from the x-ray diffraction experiments. The basic elastic properties were predicted in this study, showing that the crystalline structure of PTT is very pliable to the deformation at small strain. When the crystalline structure of PTT was stepwise deformed up to 50% of strain in chain direction under uniaxial extension condition, the change in dihedral angle of trimethylene unit from gg to tt conformation was accompanied with a large increase of stress, indicating that the phase transformation of PTT in crystalline state is difficult to occur.

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