• 제목/요약/키워드: BL52

검색결과 86건 처리시간 0.029초

C57BL/6 마우스에서 Retroviral 벡터를 이용한 Foxp3 유전자의 도입에 의한 Foxp3 단백의 발현 양상 (The expression of Foxp3 protein by retroviral vector-mediated gene transfer of Foxp3 in C57BL/6 mice)

  • 황인선;하단비;빙소진;전경익;안긴내;김대승;조진희;임재학;임신혁;황규계;지영흔
    • 대한수의학회지
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    • 제52권3호
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    • pp.183-191
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    • 2012
  • The maintenance of peripheral immune tolerance and prevention of chronic inflammation and autoimmune disease require $CD4^{+}CD25^{+}$ T cells (regulatory T cells). The transcription factor Foxp3 is essential for the development of functional, regulatory T cells, which plays a prominent role in self-tolerance. Retroviral vectors can confer high level of gene transfer and transgene expression in a variety of cell types. Here we observed that following retroviral vector-mediated gene transfer of Foxp3, transductional Foxp3 expression was increased in the liver, lung, brain, heart, muscle, spinal cord, kidney and spleen. One day after vector administration, high levels of transgene and gene expression were observed in liver and lung. At 2 days after injection, transductional Foxp3 expression level was increased in brain, heart, muscle and spinal cord, but kidney and spleen exhibited a consistent low level. This finding was inconsistent with the increase in both $CD4^{+}CD25^{+}$ T cell and $CD4^{+}Foxp3^{+}$ T cell frequencies observed in peripheral immune cells by fluorescence-activated cell-sorting (FACS) analysis. Retroviral vector-mediated gene transfer of Foxp3 did not lead to increased numbers of $CD4^{+}CD25^{+}$ T cell and $CD4^{+}Foxp3^{+}$ T cell. These results demonstrate the level and duration of transductional Foxp3 gene expression in various tissues. A better understanding of Foxp3 regulation can be useful in dissecting the cause of regulatory T cells dysfunction in several autoimmune diseases and raise the possibility of enhancing suppressive functions of regulatory T cells for therapeutic purposes.

MPEG-4 AVC|H.264 Scalable Extension을 위한 고속 모드 결정 방법 (Fast Coding Mode Decision for MPEG-4 AVC|H.264 Scalable Extension)

  • 임선희;양정엽;전병우
    • 대한전자공학회논문지SP
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    • 제45권6호
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    • pp.95-107
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    • 2008
  • 본 논문에서는 MPEG-4 AVC|H.264 SE(Scalable Extension) 부호화 복잡도의 대부분을 차지하는 모드 결정과정을 간략화시키는 시간적 및 공간적 계층 부호화에 따른 고속 모드 결정 방법을 제안한다. 우선 시간적 계층 부호화를 위해 조기스킵(Early Skip) 알고리즘과 MHM(Mode History Map)을 이용한 고속 모드 결정법을 제안한다. 조기스킵 알고리즘은 시간적으로 이전 영상과 이후 영상에 속한 참조 매크로블록의 모드만을 후보 모드로 사용하고, GOP 내에 존재하는 참조 매크로블록의 모드들로 MHM을 구성하여, 여기에 포함된 매크로블록 모드만을 후보 로드로 사용한다. 또한, 공간적 계층 부호화를 위해서는 하위 공간 계층에 대한 MHM을 구성하고, 여기에 BL_mode 만을 추가하여 상위 공간 계층의 후보 모드로 사용하는 방법을 제안한다. 제안방법은 후보 모드의 개수를 감소시킴으로써 최적 모드를 선택하기 위한 모드 결정 과정의 복잡도를 감소시킨다. 실험 결과는 제안 방법이 기존 방법에 비해 율-왜곡 성능의 큰 감소 없이 시간적 계층 부호화 방법에 대해 약 52%, 공간적 계층 부호화 방법에 대해 약 47%의 복잡도를 감소시킬 수 있음을 보여준다.

자생 산수국의 종자 발아와 유묘 생육 특성 (Characteristics of Seed Germination and Seedling Growth of Native Hydrangea serrata for. acuminata)

  • 이승연;김광진;이정식
    • 화훼연구
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    • 제16권2호
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    • pp.134-142
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    • 2008
  • 본 연구는 산수국의 종자번식에 관한 기초자료를 얻고자 수행하였다. 종자를 11월 15일, 12월 30일, 그리고 1월 23일 채취하여 파종한 결과 최종발아율이 각각 $90.0{\pm}4.16%$, $84.4{\pm}5.52%$, 그리고 $88.9{\pm}2.40%$로 휴면성이 없는 것으로 판단되었다. 광, 온도조건에 따른 발아실험 결과 최적의 발아조건은 온도는 $25^{\circ}C$이고, 광 은 명조건으로 판단되었다. 초장, 엽장, 줄기직경, 근장, 근수, 생체중, 건물중, T/R율 공히 $25/20^{\circ}C$의 주야 변온조건에서 가장 크거나 많았으나 $25^{\circ}C$ 항온조건과 큰 차이를 보이진 않았다. $15^{\circ}C$$20^{\circ}C$의 비교적 저온에서 생육이 억제되었는데, 특히 T/R율이 낮아 지상부의 생육이 더 억제된 것으로 판단되었다. 적색광은 유묘의 초장과 절간장의 신장을 촉진시켰다. 청색광 처리구에서 총엽록소, 엽록소 a/b 및 카로티노이드함량이 가장 높게 나타났고 적색+청색광 조합처리가 그 다음으로 높아 청색광이 포함된 광질 처리에서 함량이 더 높게 나타났다. 플러그 셀 수(50, 72, 128, 162, 200cell)에 따른 유묘의 생육실험결과 지상부의 생육은 50공 트레이에서 가장 좋았고 다음으로 72, 128공 순이었으며 162공과 200공 간에는 유의적인 경향이 일정하지 않았다. 대체로 셀의 부피가 큰(재식밀도가 낮은) 트레이에서 자란 유묘의 잎이 컸다.

Bacillus clausii I-52로부터 alkaline protease 유전자의 클로닝 및 발현 (Cloning and Expression of a Alkaline Protease from Bacillus clausii I-52)

  • 주한승;최장원
    • 농업생명과학연구
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    • 제45권6호
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    • pp.201-212
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    • 2011
  • 인천 연안의 심하게 오염된 갯벌로부터 강력한 세포외 알카리성 단백질 분해효소를 생산하는 호알카리성 Bacillus clausii I-52를 분리하였으며, 이 균주로부터 알카리성 단백질 분해효소의 유전자를 cloning하여 서열 분석을 하였다. Chromosome 서열이 완전히 밝혀진 Bacillus subtilis의 서열을 기초로 하여 알카리성 단백질 분해효소 및 promoter를 포함하도록 primer를 고안하여 PCR을 수행하여 2,277 bp의 DNA 단편을 얻었으며 BLAST 분석 결과 29 개의 아미노산으로 이루어진 signal peptide, 77 개의 아미노산으로 이루어진 propeptide 및 275 개의 아미노산을 갖는 활성형의 BCAP으로 구성된 총 381 개의 아미노산을 코딩하는 1,143 bp의 open reading frame을 확인하였다. 활성형 BCAP의 N-말단 아미노산은 Ala이며, 분자량 및 pI 값은 각각 27698.7 Da과 6.30으로 계산되었다. 아미노산 상동성을 분석한 결과, B. subtilis 유래의 nattokinase precursor 및 B. subtilis BSn5 유래의 subtilisin E precursor와 99%의 서열 상동성을 나타내어 B. clausii I-52 유래의 BCAP은 subtilisin 계열의 단백질 분해효소임을 확인하였다. E. coli BL21(DE3)에서 발현한 재조합 BCAP는 N-Suc-Ala-Ala-Pro-Phe-pNA 를 효율적으로 분해하였다. Refolding한 재조합 BCAP은 전형적인 serine protease inhibitor인 PMSF에 의하여 강하게 효소 활성이 억제됨으로써 serine protease 계열의 단백질 분해효소임을 알 수 있었다.

High Expression of Water-Soluble Recombinant Antigenic Domains of Toxoplasma gondii Secretory Organelles

  • Yang, Zhaoshou;Ahn, Hye-Jin;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • 제52권4호
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    • pp.367-376
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    • 2014
  • Recombinant antigenic proteins of Toxoplasma gondii are alternative source of antigens which are easily obtainable for serodiagnosis of toxoplasmosis. In this study, highly antigenic secretory organellar proteins, dense granular GRA2 and GRA3, rhoptrial ROP2, and micronemal MIC2, were analyzed by bioinformatics approach to express as water-soluble forms of antigenic domains. The transmembrane region and disorder tendency of 4 secretory proteins were predicted to clone the genes into pGEX-4T-1 vector. Recombinant plasmids were transformed into BL21 (DE3) pLysS E. coli, and GST fusion proteins were expressed with IPTG. As a result, GST fusion proteins with $GRA2_{25-105}$, $GRA3_{39-138}$, $ROP2_{324-561}$, and $MIC2_{1-284}$ domains had respectively higher value of IgG avidity. The $rGST-GRA2_{25-105}$ and $rGST-GRA3_{39-138}$ were soluble, while $rGST-ROP2_{324-561}$ and $rGST-MIC2_{1-284}$ were not. $GRA2_{31-71}$, intrinsically unstructured domain (IUD) of GRA2, was used as a linker to enhance the solubility. The $rGST-GRA2_{31-71}-ROP2_{324-561}$, a chimeric protein, appeared to be soluble. Moreover, $rGST-GRA2_{31-71}-MIC2_{1-284}$ was also soluble and had higher IgG avidity comparing to $rGST-MIC2_{1-284}$. These 4 highly expressed and water-soluble recombinant antigenic proteins may be promising candidates to improve the serodiagnosis of toxoplasmosis in addition to the major surface antigen of SAG1.

Estimation of Genetic Variance Components of Body Size Measurements in Hanwoo (Korean Cattle) Using a Multivariate Linear Model

  • Lee, Jung-Jae;Kim, Nae-Soo
    • Journal of Animal Science and Technology
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    • 제52권3호
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    • pp.167-174
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    • 2010
  • The objectives of this study were to quantify the combination values of the principal components and factors calculated using body measurements of Hanwoo (Korean Cattle) and estimate their heritabilities. The technique of multivariate analysis was used to reduce a large number of variables to a smaller number of new variables and characterize cattle according to body shape. The analyses were performed using 1,979 cattle at 12 months of age and 936 cattle at 24 months of age. The data for the analyses was obtained from progeny tests performed on Korean Cattle for 6 years from 2003 to 2008. The phenotypic correlations among these traits were estimated to range from 0.32 to 0.90 at 12 months of age and from 0.21 to 0.82 at 24 months of age. The first principal components (PC1s) indicated a weighed average of overall body measurements, accounting for 99.91% of the total variation for both periods of test. The two first PCs had positive coefficients for all body measurements. The major sources of PC, such as chest girth (CG), body length (BL), rump height (RH), and wither height (WH) were similar for both test periods. The heritabilities for PC1, the first factor score (FS1), and the second factor score (FS2) were estimated by multivariate REML method. The estimated heritabilities for PC1, FS1, and FS2 were 0.33, 0.38, and 0.40, respectively, at 12 months of age and 0.26, 0.76, and 0.58 at 24 months of age. Further studies are needed to determine whether the heritabilities of FS1 and FS2 at 24 months of age were overestimated.

Corn silk extracts did not alter the adiposity and underlying substrate utilization in high-fat diet-induced obese C57BL/6 mice

  • Kang, Eun Young;Kim, Hyun Kyung;Moyo, Knowledge Mudhibadhi;Gang, Gyoungok;Kim, Wooki;Go, Gwang-woong
    • 한국식품과학회지
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    • 제52권2호
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    • pp.144-148
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    • 2020
  • Corn silk (Okmi-su) was anciently adopted as a material for tea or beverage. Corn silk extracts (CSE) contain bioactive phytochemicals such as phenolic acid, flavonoids, ascorbic acid, tannins, and glycosides. Under the impact of these functional components, CSE has benefits for antioxidation, diuresis, anti-diabetes, and dyslipidemia recovery. Nonetheless, its role in whole-body adiposity was not investigated; therefore, the effects of CSE on obesity were evaluated in high-fat diet-induced obese mice. Mice were assigned to either group (n=12); 1) normal diet (18% kcal from fat), 2) high-fat diet (45% kcal from fat, the control), 3) high-fat diet with CSE (800 mg/kg diet), and 4) high-fat diet with orlistat (500 mg/kg diet, a comparable control for weight loss). Our results showed that body weight, adiposity, and energy expenditure in obese mice were not altered by CSE. Lean body mass tended to decrease by CSE, which can be explained by stimulation of diuresis (p=0.06). In conclusion, our results suggest that dietary consumption of CSE does not influence the adiposity and underlying substrate utilization in high-fat diet-induced obese mice.

Tissue-resident natural killer cells exacerbate tubulointerstitial fibrosis by activating transglutaminase 2 and syndecan-4 in a model of aristolochic acid-induced nephropathy

  • Wee, Yu Mee;Go, Heounjeong;Choi, Monica Young;Jung, Hey Rim;Cho, Yong Mee;Kim, Young Hoon;Han, Duck Jong;Shin, Sung
    • BMB Reports
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    • 제52권9호
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    • pp.554-559
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    • 2019
  • Despite reports suggesting that tissue-resident natural killer (trNK) cells cause ischemic kidney injury, their contribution to the development of tubulointerstitial fibrosis has not been determined. This study hypothesized that the depletion of trNK cells may ameliorate renal fibrosis by affecting transglutaminase 2/syndecan-4 interactions. Aristolochic acid nephropathy (AAN) was induced in C57BL/6 mice as an experimental model of kidney fibrosis. The mice were treated with anti-asialo GM1 (ASGM1) or anti-NK1.1 antibodies to deplete NK cells. Although both ASGM1 and NK1.1 antibodies suppressed renal $NKp46^+DX5^+$ NK cells, renal $NKp46^+DX5^-$ cells were resistant to suppression by ASGM1 or NK1.1 antibodies during the development of tubulointerstitial fibrosis in the AAN-induced mouse model. Western blot analysis showed that both antibodies increased the expression of fibronectin, transglutaminase 2, and syndecan-4. These findings indicate that trNK cells played an exacerbating role in tubulointerstitial fibrosis by activating transglutaminase 2 and syndecan-4 in the AAN-induced mouse model.

Purification and Characterization of a Recombinant Pea Chloroplastic Fructose-1, 6-bisphosphatase

  • Shin, Eun-Hye;Yoo, Yong-Cheol;Lee, Sang-Won;Hahn, Tae-Ryong
    • Journal of Applied Biological Chemistry
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    • 제44권4호
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    • pp.167-172
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    • 2001
  • A cDNA fragment encoding the chloroplastic fructose-1, 6-bisphosphatase (FBPase) was cloned via PCR from the cDNA library of pea leaves. The cloned cDNA, about 1.05 kbp without signal sequence, was introduced into a pET-28a vector for expression in E. coli strain BL21(DE3)pLysS. The recombinant FBPase was purified through $Ni^+-NTA$ affinity chromatography and characterized. Molecular mass of the monomer was about 42,000. Enzymatic activity of the purified enzyme as the native pea chloroplastic FBPase was the highest at alkaline pH (pH 9.0). The recombinant enzyme was activated by a reducing agent DTT and was insensitive to AMP. The activation energy (Ea) and Arrehenius frequency factor were 42.67 kcal/mol and $2.65{\times}10^{14}/s$, respectively, slightly higher than those of the native enzyme. $K_M$ and $V_{max}$ were $99.98{\mu}M$ and $52.9{\mu}M/min$, respectively, which were comparable with the native enzyme.

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한국인(韓國人) 중풍(中風) 환자(患者)의 Angiotensin Converting Enzyme 유전자(遺傳子) 다형성(多形成)에 관(關)한 연구(硏究) (Angiotensin Converting Enzyme(ACE) Gene Polymorphism in Korean Stroke Patients)

  • 하지영;김창환;고형균
    • Journal of Acupuncture Research
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    • 제20권2호
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    • pp.161-172
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    • 2003
  • Objective : This study was designed to investigate the relation between the angiotensin converting enzyme(ACE) gene polymorphism and stroke in the Korean population. Methods : This study was carried out on 58 stroke patients who were hospitalized in the department of acupuncture & moxibustion, college of Oriental Medicine, Kyung-Hee University and 61 healthy control subjects. Blood samples from all subjects were obtaind for DNA extration. The extracted DNA was amplified by polymerase chain reaction(PCR). PCR products were visualized by 2% agarose gel electrophoresis. Results : The sub-genotypes of ACE gene were II homozygotes, ID heterozygotes, DD homozygotes. While the distribution of ACE polymorphism in control subjects was 31%, 51%, 18%, the distribution of it in stoke patients was 33%, 52%, 16%(II, ID, DD). Thus, there was no significant different between the control and stroke groups. Conclusions : we conclude that there is no significant association between ACE gene polymorphism and storke in Korean papulation. However, the findings of this study need to be confirmed in large patients and further studies. Additional epidemiologicallly based studies of the effects and relationship between ACE or other genes and lifesyles with regard to stroke required.

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