• Title/Summary/Keyword: 형질 세포

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Selection of Chemicals for Separation of Copulated Moth of Silkworm, Bombyx mori L. (약제에 의한 가잠교미아의 할애효과에 대하여)

  • 박광의;마영일;황석조
    • Journal of Sericultural and Entomological Science
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    • v.26 no.2
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    • pp.11-15
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    • 1984
  • To save the labour reguired for separation of copulated moth during egg Production, some of the chemicals available at the market were on screen test for easy separation of copulated moth of silkworm. The obtained results are summarized as following. 1. In a separation ratio of copulated moth along with a treating time, TCTFE(Trichloro-trifluoroethane) completely separated the copulated moth in 10 minutes. The combinations of TCTFE plus Acetone (87.5 : 12.5 V/V) and TCTFE plus Acetic acid (50 : 50 V/V) take 25minutes for the complete separation of copulated moth. Use of Acetic acid solution only makes 100% separation of copulated moth in one hour and foully minutes and non$.$treatment shows only 47.5% of separation in three and half hours. On the other hand. There is no statistical significance between TCTFE Plus Acetone and control in the egg productivity. 2. The combination of TCTFE Plus Acetone (87.5 : 12.5 V/V) does not infuluence the egg productivity of the moth, showing 443 grains for an average number of egg per moth out of which 417 grains are for the number of fertilized eggs while control shows 452 grains for an avarage number of egg/moth and 428 grains for the number of fertilized eggs. However a sing1e use of TCTFE and Acetic acid shows less egg productivity and number of ferilized eggs per moth, respectively as compared to those of the control. In particular, a single use of acetic acid makes an increase of number of non-egg prodncible moth and it seems to be brought due to a chemical damage. 3. In a rearing test of the egg laid by the chemical treated moth, there are no differences among the treatments in all of the useful characters of the larvae; larval duration, survival rate, cocoon yield, single cocoon weight, single cocoon shell weight and cocoon shell ratio. In these regards, it is recognized that TCTFE can be practically used for the separation of copulated moth and the combination of TCTFE and Acetone promote its efiectiveness on the separation of copulated moth.

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Purification of cystic fluid antigen of Taenia solium metacestodes by affinity chromatography using monoclonal antibody and its antigenic characterization (단세포군항체에 의한 유구낭미충 낭액 특이항원의 순수분리 및 항원특성 관찰)

  • Kim, Suk-Il;Kang, Shin-Yong;Cho, Seung-Yull;Hwang, Eung-Soo;Cha, Chang-Yong
    • Parasites, Hosts and Diseases
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    • v.24 no.2
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    • pp.145-158
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    • 1986
  • This study was undertaken to purify cystic fluid (CF) antigen of Taenia solium metacestodes by affinity chromatogaphy using specific monoclonal antibody(McAb) and to characterize the antigenicity of the purified antigen. The hybridoma cell lines, prepared by fusion between mouse plasmacytoma and spleen cells from BALB/c mice immunized with CF, secreted antibodies reacting to various helminthic antigens. Majority of cell lines reacted to CF only but some also reacted to parenchymal antigen of T. solium metacestodes, adult T. saginata, sparganum, hydatid cystic fluid, Paragonimus westermani and Clonorchis sinensis, either in combination with CF, other antigens or independently. Cloned cells derived from monoclonal lines also produced antibodies reacting either to CF only or to other helminthes in combination or independently. These results indicated that CF of T. solium metacestodes contained proteins which possessed antigenic determinants not only specific to CF but also cross reactive with the afore-mentioned helminthes. CF of T. solium metacestodes was purified by affinity chromatography using the McAb which reacted to CF and parenchymal antigens. The affinity-purified antigen (A-Ag) and unbound pool CF (U-Ag) were separated. A-Ag showed 2 protein bands by disc-PAGE whereas CF exhibited 6 bands and U-Ag consisted of all bands CF had. The diagnostic significance of A-Ag was evaluated by ELISA in human neurocysticercosis and other helminthic and neurologic diseases. By A-Ag, the levels of the specific IgG antibody, as shown by absorbance in sera and CSF, were lower than those of CF and U-Ag. Accordingly, the sensitivity was about 70% of CF and U-Ag. However, the nonspecific positive reactions to CF and U-Ag, observed in sparganosis, T. saginata infection and paragonimiasis did not occur when A-Ag was used. These results indicated that the affinity-purified A-Ag had the higher specificity but the lower sensitivity as a diagnostic antigen in cysticercosis, probably because it only detected a single or limited numbers of monospecific antibodies among the diverse polyclonal antibodies produced in the patients with neurocysticercosis.

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Chromosome numbers of Carex section Siderostictae from Korea populations (Cyperaceae) (한국산 사초과 대사초절의 염색체 수)

  • Chung, Kyong-Sook;Yang, Jong Cheol;Lee, You-Mi
    • Korean Journal of Plant Taxonomy
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    • v.43 no.1
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    • pp.22-26
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    • 2013
  • We report somatic chromosome numbers 2n = 12 for three Carex sect. Siderostictae Franch. ex Ohwi (Cyperaceae) from Korean populations: Carex ciliatomarginata Nakai, C. okamotoi Ohwi, and C. siderosticta Hance. This study is the first chromosome number report for the species C. ciliatomarginata from Korean populations. As found in other Carex species, all the chromosomes examined in the section exhibit nonlocalized centromere (polycentric or holocentric) and large (more than ca. $1{\mu}m$ long) chromosomes. Considering the basal phylogenetic position of the section in tribe Cariceae Pax, small numbers of large chromosomes have been hypothesized as primitive characters in Cariceae, and our observation supports the hypothesis. Further investigations of chromosomes in Carex are needed for a better understanding of species richness in the genus.

Characterization of the Promoter Controling the Stage-Specific Gene Expression of Bombyx mori (누에를 이용한 시기 특이적 발현 조절 유전자 promoter 개발)

  • Park, Seung-Won;Choi, Gwang-Ho;Goo, Tae-Won;Kim, Seong-Ryul;Kang, Seok-Woo
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1466-1472
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    • 2011
  • We characterized embryo early gene (EEG)-704 promoter of the silkworm Bombyx mori, which is specifically regulated in the development stages. To determine core promoter region, 10 different partial mutant clones were tested by luciferase assay in Sf9 cells. About 1.5 kb promoter shows higher luciferase activity than constitutive promoter (BmA3). Interestingly, EEG-704 shares the same DNA sequences with BmHsp20.8 by the result of BLAST analysis; its expression is also increased under heat shock condition. Development of such promoter inducible, directly or indirectly in the developmental-stage, is very useful in making recombinant proteins in transgenic silkworms.

Production of Glucose Oxidase Using Recombinant Yeast (재조합 효모를 이용한 포도당 산화 효소의 생산)

  • 전병원;김대혁
    • KSBB Journal
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    • v.11 no.3
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    • pp.270-275
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    • 1996
  • Heterologous expression of glucose oxidase gene using recombinant yeast has been carried out. Polymerase chain reaction was conducted to obtain the gene encoding glucose oxidase from Aspergillus niger and sequence comparison indicated the cloned 1.9kb DNA fragment appeared to be the glucose oxidise structural gene containing a signal sequence for extracellular location. Transforming shuttle vector was constructed with YEp352 to express the cloned glucose oxidase gene under the control of either GAL1 or GAL10 promoter. Plate assay of recombinant yeasts has shown that GAL1 promoter was more effective in yielding glucose oxidise than GAL10 promoter. Among the five different concentrations of galactose tried, 1% galactose showed the highest induction of glucose oxidase. Cellular localization experiment of recombinant enzyme using spheroplast revealed that most of enzymes (80%) were secreted into culture media in contrast to A. niger. There is no difference in heat-stability of recombinant enzyme up to $50^{\circ}C$ compared to the glucose oxidase from A. niger However, a dramatic reduction of enzyme activity was observed in both enzymes at $60^{\circ}C$.

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Effects of Mixing Performance and Conditioned Medium on hCTLA4Ig Production in Transgenic Rice Cell Suspension Cultures (형질전환 벼 현탁세포 배양에서 혼합효율과 조정배지가 hCTLA4Ig 생산에 미치는 영향)

  • Choi, Hong-Yeol;Park, Jun-Yong;Nam, Hyung-Jin;Gong, Mi-Kyung;Yoo, Ye-Ri;Kim, Dong-Il
    • KSBB Journal
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    • v.30 no.6
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    • pp.307-312
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    • 2015
  • Transgenic rice cells using RAmy3D promoter can provide high productivity, and the production of recombinant protein is induced by sugar starvation. In this system, productivity was reduced during the scale-up processes. To ensure the influences of shear stress and oxygen transfer rate, working volume and mixing performances were investigated under various agitation speeds and working volumes. In addition, inoculation methods including suspended cells and filtered cells were compared. Working volumes and shaking speeds were 300, 450 mL and 80, 120 rpm, respectively. Hydrodynamic environment of each condition was measured numerically like mixing time and $k_La$. Good mixing performance and high shear stress were measured at high agitation speed and low volume. The highest level of hCTLA4Ig was 30.7 mg/L at 120 rpm, 300 mL. When conditioned medium was used for inoculation, increased cell growth was noticed during the day 0~4 and decreased slower than filtered cells. Compared with filtered cells, the maximum hCTLA4Ig level reached 37.8 mg/L at 120 rpm, 300 mL and lower protease activity level was observed. In conclusion mixing performance is critical factor for productivity and conditioned medium can have a positive effect on damaged cells caused by hydrodynamic shear stress.

Role of pre-C Region in the Expression and Secretion of Hepatitis B Viral Core Antigen in Yeast (효모에서 B형 간염바이러스의 내면항원의 발현과 분비에 미치는 전위내면항원의 역할)

  • 신상훈;김성기;노현모
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.1-5
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    • 1990
  • The coding sequence of hepatitis B viral core antigen (HBcAg) (subtype adr) contains two in-phase initiation codons, one for precore and the other for core antigen gene. To study the expression of core antigen and the role of precore region, the coding sequence of HBcAg with or without precore (pre-C) region were subcloned into yeast expression vector containing phosphoglycerate kinase (PGK) promoter. To study the role of upstream region in the expression of the core antigen, a series of 5' deletion mutants were also subcloned into the vector. After transformation into various host strains, the expression of HBcAg were analysed by radio-immunoassat. Under optimal condition of core antigen gene expression in yeast, the highest amount of antigen was detected in the cell line SHY4 containing pGKHBc plasmid composed of the yeast PGK gene promoter, terminator and C-gene. Regardless of the presence of precore region, core antigen was not detected in the medium but in cell extract. These results suggest that precore region cannot affect the secretion of core antigen in Saccharomyces cerevisiae.

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Immunohistochemical Observation of Plasma Cell Granuloma in Intraoral Chronic Inflammatory Lesions (구강내 만성염증병소에서 보이는 형질세포육아종의 면역조직화학적 관찰)

  • Kim, Yeon-Sook;Lee, Suk-Keun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.33 no.1
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    • pp.26-31
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    • 2011
  • Purpose: Chronic inflammatory gingival lesions occur as pyogenic granulomas or non-specific chronic suppurative lesions. Methods: Of the 59 chronic inflammatory gingival lesions examined, plasma cell granuloma (n=14), which showed an intense antibody-mediated immune reaction with the increased infiltration of plasma cells, was observed as a pseudotumor-like gingival overgrowth and myofibroblastic or fibrohistiocytitc proliferation of stromal cells with a heavy collection of plasma cells. The levels of CD3, CD20, CD31, CD68, RANKL, cathepsin G, cathepsin K, lysozyme, TNF${\alpha}$, MMP-2, and MMP-9 in the 14 cases of gingival plasma cell granuloma with immunohistochemical detection were measured to determine the pathogenetic progresses of the plasma cell granuloma compared to the common pyogenic granuloma (n=45) in the gingiva. Results: The gingival lesions of the plasma cell granuloma could be divided into three histological types, plasma cell predominant type (PPT, n=8), mixed inflammatory cell type (MICT, n=2), and sclerosed fibrosis type (SFT, n=4). The PPT showed a condensed infiltration of plasma cells into the perivascular spaces of the granulomatous lesion with frequent formation of Russel's body in their cytoplasm. The MICT showed the concomitant infiltration of many macrophages together with plasma cells, resulting in the diffuse destruction of stromal fibrous tissue. The SFT showed granulomatous lesions replaced gradually by thick collagenous fibrous tissue, resembling an inflammatory pseudotumor. The SFT expressed strongly the lymphocytic markers, CD3 and CD20, and the macrophage/monocyte markers, CD31 and CD68, but showed reduced expression of common inflammatory markers, TNF${\alpha}$, cathepsin G, lysozyme, MMP-2, and MMP-9, as well as the reduced expression of osteoclastogenic markers, RANKL and cathepsin K. Conclusion: These results suggest that a gingival plasma cell granuloma shows variable gene expression for cell-mediated immunity and stromal tissue degeneration, undergoing sclerotic fibrosis with a persistent inflammatory reaction.

Construction of Starch-Fermenting Yeast Using Protoplast Fusion Technique (원형질체 융합 방법을 이용한 전분 발효성 효모의 개발)

  • Chun, Soon-Bai;Bai, Suk;Lee, Kee-Young;Kim, Soo-Hyun;Lee, Jin-Jong;Kim, Sang-Moon
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.120-127
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    • 1990
  • Intraspecific fusion frequency between Filobasidium capsuligenum CBS6122-2 ade and met trp or met strains was $2.9\times 10^{-3}$ and $8.5\times 10^{-3}$, respectively. And intergeneric fusion frequency between Folobadidium capsuligenum CBS 4318 (cys his) and Saccharomyces cerevisiae 262-12-1 (hom3 thr1) or X2180-1A (ade thr) was $8.8\times 10^{-6}$ and $9.5\times 10^{-4}$, respectively. Nuclear fusion appears to occru in fusants between intraspecies and intergenera as strongly suggested by DNA content, nuclear staining, comparison of survival rate to UV light and mitotic segregation analysis. It was also found that $\alpha$-amylase and glucoamylase activity from intraspecific hybrids was 1.6-2.1 fold increased when compared with that from thier parents.

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Screening of Crude Drugs for the Inhibitory Effect on $NF-{\kappa}B$ Activation in Transfectant HaCaT Cells (형질전환된 각질형성세포에서 생약추출물에 의한 $NF-{\kappa}B$ 활성화 억제효과 탐색)

  • Ahn, Kwang-Seok;Kim, Seong-Kie;Moon, Ki-Young;Hahn, Bum-Soo;Kang, Sam-Sik;Kim, Yeong-Shik
    • Korean Journal of Pharmacognosy
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    • v.34 no.2 s.133
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    • pp.156-160
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    • 2003
  • $NF-{\kappa}B$ (nuclear factor-kappa B) plays a particularly central role in epidermal biology. It has been established that ultraviolet radiation (UVR) is one of the mechanisms to induce the activation of $NF-{\kappa}B$ in human skin. We previously demonstrated that melanogenic inhibitors may act through the inhibition of $NF-{\kappa}B$ activation in keratinocytes. In order to find another type of melanogenic inhibitors of $NF-{\kappa}B$ activation, various kinds of the extracts from crude drugs $(30\;{\mu}g/ml)$ were preincubated with transfectant HaCaT cells for 3 hrs and then UVR $(60\;mj/cm^2)$ was irradiated. UVR-exposed cells were incubated for another 6 hrs to measure the $NF-{\kappa}B$ activity. $NF-{\kappa}B$ activation was measured with the secreatory alkaline phosphates (SEAP) reporter gene assay using a fluorescence detection method. Among natural products, Lycium chinense, Acanthopanax senticosus, Angelica koreana, Kalopanax pictus and Asparagus cochinchinensis were the most potent inhibitors of $NF-{\kappa}B$ activation by UVR. These observations suggest that some crude drugs might act partially through the modulation of the synthesis of melanotrophic factors to decrease melanogenesis in keratinocytes.