• Tuberculosis and Respiratory Diseases
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• 제45권3호
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• pp.587-595
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• 1998

연구배경: 유전자치료에 이용되는 adenovirus를 기초로 하는 벡타는 상피세포에 강한 친화력이 있고 증식하지 않는 세포에도 유전자의 이입이 가능하며, in vivo에서 다른 백타에 비해 유전자이입 효율이 높은 장점이 있다. 그러나 adenovirus에 이해 전달된 유전자는 세포의 chromosome에 삽입되지 않기 때문에 세포가 분열하면 딸세포에 형질이 전달되지 않을 뿐 아니라 adenovirus가 숙주의 면역반응을 초래하여 이입된 유전자가 제거됨에 따라 전달된 유전자의 발현기간이 짧은 단점이 있다. 포화지방산인 butyrate는 histone의 acetylation을 증가시켜 전달된 유전자의 발현을 증가시킨다고 한다. 저자들은 유전자치료시 butyrate를 병용투여 할 경우 이입한 유전자의 발현이 증가되고, 따라서 유전자치료의 효과를 증가시킬 수 있을 것이라는 가정하에 butyrate가 herpes simplex virus thymidine kinase 유전자를 이용한 유전자치료에 미치는 영향을 조사하였다. 방 법: 악성 중피종세포인 REN세포에 Ad.CMVtk를 이입한 후 세포들을 3군으로 분류하였다. 1 군은 butyrate를 처리하지 않고 7일간 배양하였으며, 2군은 이입 후 1일째에 그리고 3군은 이입 후 2일째 1.5mM/L butyrate를 함유하는 배지에서 24시간 동안 배양한 후 butyrate가 함유되지 않은 배지로 교환하여 배양하였다. 유전자 이입 후 2일, 3일, 그리고 7일에 Western blotting을 시행하여 유전자의 발현정도를 조사하였으며, butyrate 처리 유무에 따른 bystander effect에 의한 살상효과의 차이를 조사하였다. 결 과: Butyrate를 처리하지 않은 대조군은 유전자이입 후 7일째에 유전자의 발현이 없었으나 butyrate를 처리한 군은 7일째에도 유전자의 발현이 있었다. 그리고 butyrate를 처리한 경우 bystander effect에 의한 살상효과가 유의하게 증가되었다. 결 론: 이상의 결과로 butyrate는 유전자의 발현을 증가시킴에 의해 bystander effect를 증가시킴을 알 수 있었으며, 향 후 adenovirus를 이장한 유전자치료의 효과를 증가시킬 수 있는 새로운 방법이 될 수 있을 것으로 생각된다.

• 지능정보연구
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• 제27권3호
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• pp.157-173
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• 2021

소비자의 욕구와 관심에 맞추어 개인화된 제품을 추천하는 추천 시스템은 비즈니스에 필수적인 기술로서의 그 중요성이 증가하고 있다. 추천 시스템의 대표적인 모형 중 협업 필터링은 우수한 성능으로 다양한 분야에서 활용되고 있다. 그러나 협업필터링은 사용자-아이템의 선호도 정보가 충분하지 않을 경우 성능이 저하되는 희소성의 문제가 있다. 또한 실제 평점 데이터의 경우 대부분 높은 점수에 데이터가 편향되어 있어 심한 불균형을 갖는다. 불균형 데이터에 협업 필터링을 적용할 경우 편향된 클래스에 과도하게 학습되어 추천 성능이 저하된다. 이러한 문제를 해결하기 위해 많은 선행연구들이 진행되어 왔지만 추가적인 외부 데이터 또는 기존의 전통적인 오버샘플링 기법에 의존한 추천을 시도하였기에 유용성이 떨어지고 추천 성능 측면에서 한계점이 있었다. 본 연구에서는 CGAN을 기반으로 협업 필터링 구현 시 발생하는 희소성 문제를 해결함과 동시에 실제 데이터에서 발생하는 데이터 불균형을 완화하여 추천의 성능을 높이는 것을 목표로 한다. CGAN을 이용하여 비어있는 사용자-아이템 매트릭스에 실제와 흡사한 가상의 데이터를 생성하여, 희소성을 가지고 있는 기존의 매트릭스로만 학습한 것과 비교했을 때 높은 정확도가 예상된다. 이 과정에서 Condition vector y를 이용하여 소수 클래스에 대한 분포를 파악하고 그 특징을 반영하여 데이터를 생성하였다. 이후 협업 필터링을 적용하고, 하이퍼파라미터 튜닝을 통해 추천 시스템의 성능을 최대화하는데 기여하였다. 비교 대상으로는 전통적인 오버샘플링 기법인 SMOTE, BorderlineSMOTE, SVM-SMOTE, ADASYN와 GAN을 사용하였다. 결과적으로 데이터 희소성을 가지고 있는 기존의 실제 데이터뿐만 아니라 기존 오버샘플링 기법들보다 제안 모형의 추천 성능이 우수함을 확인하였으며, RMSE, MAE 평가 척도에서 가장 높은 예측 정확도를 나타낸다는 사실을 증명하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제23권3호
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• pp.292-301
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• 2010

Follicle-stimulating hormone (FSH) is a pituitary glycoprotein hormone that is encoded by separate alpha- and betasubunit genes. It plays a key role in stimulating and regulating ovarian follicular development and egg production in chicken. FSH signal transduction is mediated by the FSH receptor (FSHR) that exclusively interacts with the beta-subunit of FSH, but characterization of prokaryotic expression of the FSHb gene and its effect on the expression of the FSHR gene in local chickens have received very little attention. In the current study, the cDNA fragment of the FSHb gene from Dagu chicken was amplified using reverse transcription polymerase chain reaction (RT-PCR), and inserted into the pET-28a (+) vector to construct the pET-28a-FSHb plasmid. After expression of the plasmid in E. coli BL21 (DE3) under inducing conditions, the recombination protein, $FSH{\beta}$ subunit, was purified and injected into the experimental hens and the effect on the mRNA expression levels of the FSHR gene was investigated. Sequence comparison showed that the coding region of the FSHb gene in the local chicken shared 99%-100% homology to published nucleotides in chickens; only one synonymous nucleotide substitution was detected in the region. The encoded amino acids were completely identical with the reported sequence, which confirmed that the sequences of the chicken FSHb gene and the peptides of the $FSH{\beta}$ subunit are highly conserved. This may be due to the critical role of the normal function of the FSHb gene in hormonal specificity and regulation of reproduction. The results of gene expression revealed that a recombinant protein with a molecular weight of about 19 kDa was efficiently expressed and it was identified by Western blotting analysis. After administration of the purified $FSH{\beta}$ protein, significantly higher expression levels were demonstrated in uterus, ovary and oviduct samples (p<0.05). These observations suggested that the expressed $FSH{\beta}$ protein possesses biological activity, and has a potential role in regulation of reproductive physiology in chickens.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권4호
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• pp.378-384
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• 2005

Vanillin is one of the world's principal flavoring compounds, and is used extensively in the food industry. The potential vanillin production of the bacteria was compared to select and clone genes which were appropriate for highly productive vanillin production by E. coli. The fcs (feruloyl-CoA synthetase) and ech (enoyl-CoA hydratase/aldolase) genes cloned from Amycolatopsis sp. strain HR104 and Delftia acidovorans were introduced to pBAD24 vector with $P_{BAD}$ promoter and were named pDAHEF and pDDAEF, respectively. We observed 160 mg/L vanillin production with E. coli harboring pDAHEF, whereas 10 mg/L of vanillin was observed with pDAHEF. Vanillin production was optimized with E. coli harboring pDAHEF. Induction of the fcs and ech genes from pDAHEF was optimized with the addition of 13.3 mM arabinose at 18 h of culture, from which 450 mg/L of vanillin was produced. The feeding time and concentration of ferulic acid were also optimized by the supplementation of $0.2\%$ ferulic acid at 18 h of culture, from which 500 mg/L of vanillin was obtained. Under the above optimized condition of arabinose induction and ferulic acid supplementation, vanillin production was carried out with four different types of media, M9, LB, 2YT, and TB. The highest vanillin production, 580 mg/L, was obtained with LB medium, a 3.6 fold increase in comparison to the 160 mg/L obtained before the optimization of vanillin production.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.40-44
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• 2003

Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

• BMB Reports
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• 제37권3호
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• pp.282-291
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• 2004

Phytases catalyze the release of phosphate from phytic acid. Phytase-producing microorganisms were selected by culturing the soil extracts on agar plates containing phytic acid. Two hundred colonies that exhibited potential phytase activity were selected for further study. The colony showing the highest phytase activity was identified as Aspergillus niger and designated strain 113. The phytase gene from A. niger 113 (phyI1) was isolated, cloned, and characterized. The nucleotide and deduced amino acid sequence identity between phyI1 and phyA from NRRL3135 were 90% and 98%, respectively. The identity between phyI1 and phyA from SK-57 was 89% and 96%. A synthetic phytase gene, phyI1s, was synthesized by successive PCR and transformed into the yeast expression vector carrying a signal peptide that was designed and synthesized using P. pastoris biased codon. For the phytase expression and secretion, the construct was integrated into the genome of P. pastoris by homologous recombination. Over-expressing strains were selected and fermented. It was discovered that ~4.2 g phytase could be purified from one liter of culture fluid. The activity of the resulting phytase was 9.5 U/mg. Due to the heavy glycosylation, the expressed phytase varied in size (120, 95, 85, and 64 kDa), but could be deglycosylated to a homogeneous 64 kDa species. An enzymatic kinetics analysis showed that the phytase had two pH optima (pH 2.0 and pH 5.0) and an optimum temperature of $60^{\circ}C$.

• 통상정보연구
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• 제13권3호
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• pp.77-95
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• 2011

본고에서는 무역보험과 환위험이 우리나라의 수출에 미치는 영향을 분석하였다. 먼저 공적분 검정을 통해 적어도 하나의 공적분 벡터가 존재하는 것을 확인하였고, CCR, DOLS, FMOLS와 같은 공적분 벡터식과 오차수정모형을 추정하였다. 설정된 모형 내 변수들이 수출에 미치는 영향의 방향이 무역보험, 세계경기, 상대가격, 는 양(+)으로, 환위험과 실업률은 음(-)으로 나타났다. 또한 모형1의 오차수정 모형의 경우 단기 불균형에서 균형으로의 조정역할은 세계경기와 상대가격이 하고 있으며, 모형2의 오차수정모형의 경우 단기 불균형에서 균형으로의 조정역할은 환율변동성이 수행하고 있는 것으로 분석되었다. 다음으로 충격반응분석 결과 모형 1과 모형 2에서 무역보험 충격에 대한 수출의 반응은 충격을 받은 시점에서 2개월까지는 상승하고, 이후에는 상승세가 둔화되면서 상승효과가 소멸되는 것으로 나타났다. 모형 2에서 환율변동성 충격에 대한 수출의 반응은 충격을 받은 시점에서 4개월이 되는 시점에서 가장 큰 폭으로 하락하고 빠른 속도로 감소하고 있음을 알 수 있었다. 결과적으로 우리나라의 경우 무역보험은 수출촉진을 위한 무역정책의 역할을 수행하며, 환율변동성 확대는 환위험을 증가시켜 수출을 위축시키는 것으로 나타났다. 따라서 무역보험의 정책적 지원이 확대되어야 하며, 외환시장의 다변화와 거래 규모 확대가 필요하다.

• BMB Reports
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• 제39권3호
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• pp.253-262
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• 2006

Parkinson's disease (PD) is a common neurodegenerative disorder and is characterized by the progressive loss of dopaminergic neurons in the substantia nigra. Although many studies showed that the aggregation of $\alpha$-synuclein might be involved in the pathogenesis of PD, its protective properties against oxidative stress remain to be elucidated. In this study, human wild type and mutant $\alpha$-synuclein genes were fused with a gene fragment encoding the nine amino acid trans activator of transcription (Tat) protein transduction domain of HIV-l in a bacterial expression vector to produce a genetic in-frame WT Tat-$\alpha$-synuclein (wild type) and mutant Tat-a-synucleins (mutants; A30P and A53T), respectively, and we investigated the protective effects of wild type and mutant Tat-$\alpha$-synucleins in vitro and in vivo. WT Tat-$\alpha$-synuclein rapidly transduced into an astrocyte cells and protected the cells against paraquat induced cell death. However, mutant Tat-$\alpha$-synucleins did not protect at all. In the mice models exposed to the herbicide paraquat, the WT Tat-$\alpha$-synuclein completely protected against dopaminergic neuronal cell death, whereas mutants failed in protecting against oxidative stress. We found that these protective effects were characterized by increasing the expression level of heat shock protein 70 (HSP70) in the neuronal cells and this expression level was dependent on the concentration of transduced WT Tat-$\alpha$-synuclein. These results suggest that transduced Tat-$\alpha$-synuclein might protect cell death from oxidative stress by increasing the expression level of HSP70 in vitro and in vivo and this may be of potential therapeutic benefit in the pathogenesis of PD.

• Journal of Microbiology and Biotechnology
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• 제28권9호
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• pp.1547-1553
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• 2018

Hyaluronidases are a family of enzymes that catalyse the breakdown of hyaluronic acid, which is abundant in the extracellular matrix and cumulus oocyte complex. To investigate the activity of recombinant bovine sperm hyaluronidase 1 (SPAM1) and determine the effect of the Asn-X-Ser/Thr motif on its activity, the bovine SPAM1 open reading frame was cloned into the mammalian expression vector pCXN2 and then transfected to the HEK293 cell line. Expression of recombinant bovine hyaluronidase was estimated using a hyaluronidase activity assay with gel electrophoresis. Recombinant hyaluronidase could resolve highly polymeric hyaluronic acid and also caused dispersal of the cumulus cell layer. Comparative analysis with respect to enzyme activity was carried out for the glycosylated and deglycosylated bovine sperm hyaluronidase by N-glycosidase F treatment. Finally, mutagenesis analysis revealed that among the five potential N-linked glycosylation sites, only three contributed to significant inhibition of hyaluronic activity. Recombinant bovine SPAM1 has hyaluronan degradation and cumulus oocyte complex dispersion ability, and the N-linked oligosaccharides are important for enzyme activity, providing a foundation for the commercialization of hyaluronidase.

• 한국산학기술학회논문지
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• 제18권1호
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• pp.688-695
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• 2017

본 논문은 파력 에너지 수집 장치에 사용할 수 있는 영구자석 선형 동기발전기의 특성 해석에 관한 것이다. 파력 에너지는 요요시스템과 같은 기구로 부터 얻어진다. 영구자석을 이용한 선형 발전기는 영구자석의 자력을 통해 별도의 전원공급이 필요 없고 유지 보수가 간단한 장점을 가지고 있다. 또한 높은 에너지 밀도를 갖는 희토류의 사용으로 영구자석 기기는 소형화 및 경량화가 가능하며 보다 높은 에너지 변환 효율을 얻을 수 있다. 영구자석 선형 동기발전기 특성 해석을 위해 2차원 극 좌표계 및 자기 벡터 포텐셜에 근거하여 영구자석과 전기자 반작용 자계해석을 수행 하였다. 해석 해를 이용하여 정현적인 속도입력에 의해 유도되는 유기기전력의 특성 식을 유도하고, 동일한 방법으로 역기전력 상수, 저항, 자기인덕턴스와 상호인덕턴스와 같은 전기적 파라미터를 얻었다. 본 논문에서 사용한 공간고조파법의 결과는 2차원 유한요소해석법 결과와 비교하여 잘 일치하는 것을 확인하였다. 이 결과는 영구자석 형 선형 발전기의 특성을 이해하는 것과 해석방법의 비교연구, 설계 최적화, 그리고 기기의 동적 모델링에 기여할 수 있다.