• Title/Summary/Keyword: GPD1

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Production of a Functional Mouse Interferon ${\gamma}$from Recombinant Saccharomyces cerevisiae

  • Lim, Young-Yi;Park, Seung-Moon;Jang, Yong-Suk;Yang, Moon-Sik;Kim, Dae-Hyuk
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.537-543
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    • 2003
  • The mouse interferon gene (MuIFN-${\gamma}$) was cloned and then used to transform Saccharomyces cerevisiae. Expressed MuIFN-$\{gamma}$ protein (MuIFN-${\gamma}$) was successfully secreted into culture medium due to the presence oi the signal peptide of rice amylase 1A. Two different promoters fused to MuIFN-${\gamma}$ were tested: glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter and a yeast hybrid ADH2-GPD (AG) promoter consisting of alcohol dehydrogenase II (ADH2) and GPD promoter. Using the hybrid promoter, the accumulation of MuIFN-${\gamma}$transcript was the highest after the 24 h cultivation, and then gradually decreased as the cultivation proceeded. However, both cell growth and recombinant MuIFN-${\gamma}$production reached their peaks after the 4-day cultivation. It was possible to produce 6.5 mg/l of MuIFN-${\gamma}$ without any changes in cell growth. Using GPD promoter, the MuIFN-${\gamma}$ transcript accumulation and the recombinant MuIFN-${\gamma}$ production followed the same pattern as the cell growth. However. compared to that of the hybrid promoter, the production of recombinant MuIFN-${\gamma}$ was 0.2 mg/l. The secreted MuIFN-${\gamma}$ had estimated molecular masses of 21 kDa and 23 kDa, which were larger than that of the encoded size due to glycosylation. The protection assay against the viral infection indicated that the recombinant MuIFN-${\gamma}$ was bioactive.

Heterologous Expression of Human Ferritin H-chain and L-chain Genes in Saccharomyces cerevisiae (재조합 효모를 이용한 사람 H-Chain 교 L-Chain Ferritin의 생산)

  • 서향임;전은순;정윤조;김경숙
    • KSBB Journal
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    • v.17 no.2
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    • pp.162-168
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    • 2002
  • Human ferritin H- and L-chain genes(hfH and hfL) were cloned into the yeast shuttle vector YEp352 with various promoters, and the vectors constructed were used to transform Saccharomyces cerevisiae 2805. Three different promoters fused to hfH and hfL were used: galactokinase 1 (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase(GPD) promoter and alcohol dehydrogenase 1(ADH1 ) promoter. SDS-polyacrylamide gel electrophoresis and Western blotting analyses displayed expression of the introduced hfH and hfL. In the production of both ferritin H and L subunits GAL1 promoter was more effective than GPD promoter or ADH1 promoter. Ferritin H and L subunits produced in S. cerevisiae were spontaneously assembled into its holoproteins as proven on native polyacrylamide gels. Both recombinant H and L-chain ferritins were catalytically active in forming iron core. When the cells were cultured in the medium containing 10 mM ferric citrate, the cell-associated concentration of iron was 174.9 $\mu\textrm{g}$ Per gram(dry cell weight) for the recombinant yeast YG-L and 148.8 $\mu\textrm{g}$ Per gram(dry cell weight) for the recombinant yeast YG-L but was 49.4 $\mu\textrm{g}$ Per gram(dry cell weight) in the wild type, indicating that the iron contents of yeast is improved by heterologous expression of human ferritin H-chain or L-chain genes.

Enhanced and Targeted Expression of Fungal Phytase in Saccharomyces cerevisiae

  • LIM, YOUNG-YI;EUN-HA PARK;JI-HYE KIM;SEUNG-MOON PARK;HYO-SANG JANG;YOUN-JE PARK;SEWANG YOON;MOON-SIK YANG;DAE-HYUK KIM
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.915-921
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    • 2001
  • Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. In order to express a high level of fungal phytase in Saccharomyces cerevisiae, various expression vectors were constructed with different combinations of promoters, translation enhancers, signal peptides, and terminator. Three different promoters fused to the phytase gene (phyA) from Aspergillus niger were tested: a galactokinase (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, and yeast hybrid ADH2-GPD promoter consisting of alcohol dehydrogenase II (ADH2) and a GPD promoter. The signal peptides of phytase, glucose oxidase (GO), and rice amylase 1A(RAmy1A) were included. Plus, the translation enhancers of the ${\Omega}$ sequence and UTR70 from the tobacco mosaic virus (TMV) and spinach, respectively, were also tested. Among the recombinant vectors, pGphyA06 containing the GPD promoter, the ${\Omega}$ sequence, RAmy1A, and GAL7 terminator expressed the highest phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase was also performed by inserting an endoplasmic reticulum (ER) retention signal, KDEL sequence, into the C-terminus of the phytase within the vector pHphyA-6. It appeared that the KDEL sequence directed most of the early expression of phytase into the intracellular compartment yet more than $60\%$ of the total phytase activity was still retained within the cell even after the prolonged (>3 days) incubation of the transformant. However, the intracellular enzyme activity of the transformant without a KDEL sequence was as high as that of the extracellular one, thereby strongly suggesting that the secretion of phytase in S. cerevisiae appeared to be the rate-limiting step for the expression of a large amount of extracellular recombinant phytase, when compared with other yeasts.

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Discriminative Training of Predictive Neural Network Models (예측신경회로망 모델의 변별력 있는 학습)

  • Na, Kyung-Min;Rheem, Jae-Yeol;Ann, Sou-Guil
    • The Journal of the Acoustical Society of Korea
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    • v.13 no.1E
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    • pp.64-70
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    • 1994
  • Predictive neural network models are powerful speech recognition models based on a nonlinear pattern prediction. But those models suffer from poor discrimination between acoustically similar words. In this paper we propose an discriminative training algorithm for predictive neural network models. This algorithm is derived from GPD (Generalized Probabilistic Descent) algorithm coupled with MCEF(Minimum Classification Error Formulation). It allows direct minimization of a recognition error rate. Evaluation of our training algoritym on ten Korean digits shows its effectiveness by 30% reduction of recognition error.

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Effect of Ginseng Saponins on Phagocytosis of Feline Peripheral Blood Phagocytes (고양이 말초혈액 탐식세포의 탐식능에 있어서 인삼 사포닌의 효과)

  • 양만표;박세형;윤영원
    • Journal of Veterinary Clinics
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    • v.15 no.1
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    • pp.116-123
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    • 1998
  • 고양이 말초혈액 탐식세포(단핵구세포(MNC) 및 다형핵백혈구(PMNC))의 탐식능 에 있어서 인삼 saponin(ginseng total saponin(G75), ginseng PT saponin(GPT) 및 ginseng PD saponin(GPD))의 면역증강 효과를 flow cytometry를 이용하여 분석하였다. 인삼 ssponins을 직접 첨가하여 배양한 MNC 및 PMNC에서는 탐식중강 효과가 나타나지 않았다. 각각의 인삼 saponin을 첨가하여 배양한 PMNC 및 MNC 배양상충액의 존재하에 PMNC 및 MNC의 탐식능을 겅토한 결과, MNC의 탐식능은 Gff 첨가 PMNC 배양상충액과 GTS 및 GPT 첨가 MNC 배양상충액의 존재하에서 약간의 탐식증강 효과를 보였다. PMNC 탐식능의 경우에는 GPD 첨가 PMNC 배양상충액에서 미약한 탐식증강 효과가 나타났으나, 각각의 인 삼 saponin 첨가 MNC 배양상충액 존재하에서는 모두 현저한 탐식중강 효과를 나타내었다. 이상의 결과로부터 고양이 말초혈액 탐식세포의 탐식증강 효과는 인삼 saponin의 직접적인 작용보다는 인삼 saponin에 의해 활성화된 단핵구세포에서 분비되는 가용성물질에 의해 단 핵구세포보다는 다형핵백혈구에서 현저하게 탐식효과가 증강되는 것으로 판단되었다.

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Analysis of torrential rainfall characteristics using 'zero-inflated models' ('0-과잉 모형'을 이용한 집중호우의 발생특성 분석)

  • Kim, Sang Ug
    • Proceedings of the Korea Water Resources Association Conference
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    • 2017.05a
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    • pp.453-453
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    • 2017
  • 본 연구에서는 최근 기후변화로 인한 집중호우의 발생횟수의 경향을 확률적으로 분석함에 있어 1개월 동안 80 mm/day 이상의 강우사상을 집중호우로 정의하여, 대구 및 부산 강우관측소로부터 수집된 384개월 동안의 집중호우를 분석하였다. 집중호우 월별 발생횟수와 같은 형식의 자료의 확률적 분석은 대개 Poisson 분포 (POI)가 사용되나 자료에 포함된 0자료의 과잉은 확률분포를 왜곡시키는 문제를 발생시킨다. 본 연구에서는 이 문제를 개선하기 위하여 개발된 일반화 Poisson 확률분포 (GPD), 0-과잉 Poisson 확률분포 (ZIP), 0-과잉 일반화 Poisson 확률분포 (ZIGP), Bayesian 0-과잉 일반화 Poisson 확률분포 (Bayesian ZIGP)를 집중호우 자료에 적용하고, 5개 모형의 특성을 비교분석하였으며, Bayesian ZIGP 모형의 구축에 있어서는 정보적 사전분포를 사용함으로써 모형의 정확도를 개선하였다. 분석결과 분석하고자 하는 자료에 0이 과다하게 포함되어 있는 경우 POI 및 GPD 분포는 관측결과와는 다른 결과를 제시하여 적절한 모형으로 고려되지 못함을 알 수 있었다. 5가지 모형 중 정보적 사전분포를 탑재한 Bayesian ZIGP 모형이 가장 관측 자료와 유사한 결과를 도출하였으나 모형의 구축에 수반되는 실용적인 측면을 고려하면 ZIP 모형도 충분히 사용될 수 있는 모형으로 추천되었다.

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Estimation of Car Insurance Loss Ratio Using the Peaks over Threshold Method (POT방법론을 이용한 자동차보험 손해율 추정)

  • Kim, S.Y.;Song, J.
    • The Korean Journal of Applied Statistics
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    • v.25 no.1
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    • pp.101-114
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    • 2012
  • In car insurance, the loss ratio is the ratio of total losses paid out in claims divided by the total earned premiums. In order to minimize the loss to the insurance company, estimating extreme quantiles of loss ratio distribution is necessary because the loss ratio has essential prot and loss information. Like other types of insurance related datasets, the distribution of the loss ratio has heavy-tailed distribution. The Peaks over Threshold(POT) and the Hill estimator are commonly used to estimate extreme quantiles for heavy-tailed distribution. This article compares and analyzes the performances of various kinds of parameter estimating methods by using a simulation and the real loss ratio of car insurance data. In addition, we estimate extreme quantiles using the Hill estimator. As a result, the simulation and the loss ratio data applications demonstrate that the POT method estimates quantiles more accurately than the Hill estimation method in most cases. Moreover, MLE, Zhang, NLS-2 methods show the best performances among the methods of the GPD parameters estimation.

Expression of Nutritionally Well-balanced Protein, AmA1, in Saccharomyces cerevisiae

  • Kim, Tae-Geum;Kim, Ju;Kim, Dae-Hyuk;Yang, Moon-Sik
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.3
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    • pp.173-178
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    • 2001
  • Food yeast, Saccharomyces cerevisiae, is a safe organism with a long history of use for the production of biomass rich in high quality proteins and vitamins. AmA1, a seed storage albumin from Amaranthus hypochondriacus, has a well-balanced amino acid composition and high levels of essential amino acids and offers the possibility of further improving food animal feed additives. In order to find an effective means of expressing AmA1 in yeast, the gene was cloned into an episomal shuttle vector. Four different promoters were tested: the glyceraldehyde-3-phosphate dehydrogenase promoter, galactose dehydrogenase 10 promoter, alcohol dehydrogenase II promoter, and a hybrid ADH2-GPD promoter. The recombinant AmA1 genes were then introduced into the yeast Saccharomyces cerevisiae 2805. Northern and Western blot analyses of the yeast under appropriate conditions revealed that AmA1 was expressed by all four promoters at varying levels. An enzyme-linked immunosorbent assay demonstrated that the amount of AmA1 protein in the recombinant yeast was 1.3-4.3% of the total soluble proteins. The highest expression level was obtained from the hybrid ADH2-GPD promoter.

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The Irradiated Lung Volume in Tangential Fields for the Treatment of a Breast (유방암의 접선 조사시 피폭 폐용적)

  • Oh Young Taek;Kim Juree;Kang Haejin;Sohn Jeong Hye;Kang Seung Hee;Chun Mison
    • Radiation Oncology Journal
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    • v.15 no.2
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    • pp.137-143
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    • 1997
  • Purpose : Radiation pneumonitis is one of the complications caused by radiation therapy that includes a Portion of the lung tissue. The severity of radiation induced pulmonary dysfunction depends on the irradiated lung volume, total dose, dose rate and underlying Pulmonary function. It also depends on whether chemotherapy is done or not. The irradiated lung volume is the most important factor to predict the pulmonary dysfunction in breast cancer Patients following radiation therapy. There are some data that show the irradiated lung volume measured from CT scans as a part of treatment Planning with the tangential beams. But such data have not been reported in Korea. We planned to evaluate the irradiated lung volume quantitatively using CT scans for the breast tangential field and search for useful factors that could Predict the irradiated lung volume Materials and Methods : The lung volume was measured for 25 patients with breast cancer irradiated with tangential field from Jan.1995 to Aug.1996. Parameters that can predict the irradiated lung volume included; (1) the peruendicular distance from the Posterior tangential edge to the posterior part of the anterior chest wall at the center of the field (CLD) ; (2) the maximum perpendicular distance from the posterior tangential field edge to the posterior Part of the anterior chest wall (MLD) ; (3) the greatest perpendicular distance from the Posterior tangential edge to the posterior part of anterior chest wall on CT image at the center of the longitudinal field (GPD) ; (4) the length of the longitudinal field (L). The irradiated lung volume(RV), the entire both lung volume(EV) and the ipsilateral lung volume(IV) were measured using dose volume histogram. The relationship between the irradiated lung volume and predictors was evaluated by regression analysis. Results :The RV is 61-279cc (mean 170cc), the RV/EV is $2.9-13.0\%\;(mean\;5.8\%)$ and the RV/IV is $4.9-29.0\%\;(mean\;12.2\%)$. The CLD, the MLD and the GPD ave 1.9-3.3cm, 1.9-3.3cm and 1.4-3.1cm respectively. The significant relations between the irradiated lung volume such as RV. RV/EV, RV/IV and parameters such as CLD, MLD, GPO, L. $CLD\timesL,\;MLD\timesL\;and\;GPD\timesL$ are not found with little variances in parameters. The RV/IV of the left breast irradiation is significantly larger than that of the right but the RV/EVS do not show the differences. There is no symptomatic radiation pneumonitis at least during 6 months follow up. Conclusion : The significant relationship between the irradiated lung volume and predictors is not found with little variation on parameters. The irradiated lung volume in the tangential held is liss than $10\%$ of entire lung volume when CLO is less than 3cm. The RV/IV of the left tangential field is larger than that of the right but there was no significant differences in RV/EVS. Symptomatic radiation pneumonitis has not occurred during minimum 6 months follow up.

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Comparison of probability distributions to analyze the number of occurrence of torrential rainfall events (집중호우사상의 발생횟수 분석을 위한 확률분포의 비교)

  • Kim, Sang Ug;Kim, Hyeung Bae
    • Journal of Korea Water Resources Association
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    • v.49 no.6
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    • pp.481-493
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    • 2016
  • The statistical analysis to the torrential rainfall data that is defined as a rainfall amount more than 80 mm/day is performed with Daegu and Busan rainfall data which is collected during 384 months. The number of occurrence of the torrential rainfall events can be simulated usually using Poisson distribution. However, the Poisson distribution can be frequently failed to simulate the statistical characteristics of the observed value when the observed data is zero-inflated. Therefore, in this study, Generalized Poisson distribution (GPD), Zero-Inflated Poisson distribution (ZIP), Zero-Inflated Generalized Poisson distribution (ZIGP), and Bayesian ZIGP model were used to resolve the zero-inflated problem in the torrential rainfall data. Especially, in Bayesian ZIGP model, a informative prior distribution was used to increase the accuracy of that model. Finally, it was suggested that POI and GPD model should be discouraged to fit the frequency of the torrential rainfall data. Also, Bayesian ZIGP model using informative prior provided the most accurate results. Additionally, it was recommended that ZIP model could be alternative choice on the practical aspect since the Bayesian approach of this study was considerably complex.