• 제목/요약/키워드: Replication protein A

검색결과 326건 처리시간 0.027초

한탄바이러스 Nucleocapsid Protein 발현에 있어 S Genome 내 Noncoding Region의 역할 (The Role of Noncoding Region in Hantaan Viral S Genome for Expression of Nucleocapsid Protein)

  • 유정희;이연승;이호동;박찬;박근용;이평우
    • 대한바이러스학회지
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    • 제30권1호
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    • pp.39-49
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    • 2000
  • The genome of Hantaan virus, the prototype of the hantavirus genus, is composed of three segmented, single stranded negative sense RNA genome. The 5' and 3' termini of the Hantaan virus RNA genome contain noncoding regions (NCRs) that are highly conserved and complementary to form panhandle structures. There are some reports that these NCRs seems to control gene expression and viral replication in influenza virus and vesicular stomatitis virus. In this study, we examined whether NCRs in Hantaan virus playa role in expression of the viral nucleocapsid protein (Np) and foreign (luciferase) gene. The 5' and/or 3' NCR-deleted mutants were constructed and analysed. The Np expression of 5' NCR-deleted clone was similar to that of the clone containing full S genome. In the case of 3' NCR-deleted clone, it showed 40% reduction. To investigate the role of NCR in foreign gene expression, the clones which are replaced ORF of Hantaan viral Np gene with that of luciferase gene were constructed. The results were similar to those of the experiments using Np gene. These results suggest that 3' NCR is more important than 5' NCR in protein expression. To find out a critical region of 3' NCR in protein expression, several clones with a deleted part of 3' NCR were constructed and analyzed. The deletion of the conserved region in 3' NCR showed $20{\sim}30%$ decrease in Np expression. However there were no change in luciferase activities between clones with or without non-conserved region of 3' NCR. These results suggest that the 3' NCR of Hantaan virus S genome, especially conserved region in 3' NCR, plays an important role in the expression of Hantaan viral Np and foreign genes.

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Possible Mechanism Underlying the Antiherpetic Activity of a Proteoglycan Isolated from the Mycelia of Ganoderma lucidum in Vitro

  • Li, Zubing;Liu, Jing;Zhao, Yifang
    • BMB Reports
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    • 제38권1호
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    • pp.34-40
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    • 2005
  • GLPG (Ganoderma lucidum proteoglycan) was a bioactive fraction obtained by the liquid fermentation of the mycelia of Ganoderma lucidum, EtOH precipitation, and DEAE-cellulose column chromatography. GLPG was a proteoglycan with a carbohydrate: protein ratio of 10.4: 1. Its antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) were investigated using a cytopathic inhibition assay. GLPG inhibited cell death in a dose-dependent manner in HSV-infected cells. In addition, it had no cytotoxic effect even at 2 mg/ml. In order to study the mode of action of the antiviral activity of GLPG, cells were treated with GLPG before, during, and after infection, and viral titer in the supernatant of cell culture 48 h post-infection was determined using a $TCID_{50}$ assay. The antiviral effects of GLPG were more remarkable before viral treatment than after treatment. Although the precise mechanism has yet to be defined, our work suggests that GLPG inhibits viral replication by interfering with the early events of viral adsorption and entry into target cells. Thus, this proteoglycan appears to be a candidate anti-HSV agent.

Construction of an Escherichia-Pseudomonas Shuttle Vector Containing an Aminoglycoside Phosphotransferase Gene and a lacZ' Gene for $\alpha$-Complementation

  • Lee, Bheong-Uk;Hong, Ja-Heon;Kahng, Hyung-Yeel;Oh, Kye-Heon
    • Journal of Microbiology
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    • 제44권6호
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    • pp.671-673
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    • 2006
  • A new 4.87 kb Escherichia-Pseudomonas shuttle vector has been constructed by inserting a 1.27 kb DNA fragment with a replication origin of a Pseudomonas plasmid pRO1614 into the 3.6 kb E. coli plasmid pBGS18. This vector, designated pJH1, contains an aminogly-coside phosphotransferase gene (aph) from Tn903, a lacZ' gene for $\alpha$-complementation and a versatile multiple cloning site possessing unique restriction sites for EcoRI, SacI, KpnI, SmaI, BamHI, XbaI, SalI, BspMI, PstI, SphI, and HindIII. When pJH1 was transformed into E. coli DHS${\alpha}$ and into P. putida HK-6, it was episomally and stably maintained in both strains. In addition, the enhanced green fluorescent protein (EGFP) gene which was transcriptionally cloned into pJH1 rendered E. coli cells fluorescence when its transformants were illuminated at 488 nm.

Replication and packaging of Turnip yellow mosaic virus RNA containing Flock house virus RNA1 sequence

  • Kim, Hui-Bae;Kim, Do-Yeong;Cho, Tae-Ju
    • BMB Reports
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    • 제47권6호
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    • pp.330-335
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    • 2014
  • Turnip yellow mosaic virus (TYMV) is a spherical plant virus that has a single 6.3 kb positive strand RNA as a genome. In this study, RNA1 sequence of Flock house virus (FHV) was inserted into the TYMV genome to test whether TYMV can accommodate and express another viral entity. In the resulting construct, designated TY-FHV, the FHV RNA1 sequence was expressed as a TYMV subgenomic RNA. Northern analysis of the Nicotiana benthamiana leaves agroinfiltrated with the TY-FHV showed that both genomic and subgenomic FHV RNAs were abundantly produced. This indicates that the FHV RNA1 sequence was correctly expressed and translated to produce a functional FHV replicase. Although these FHV RNAs were not encapsidated, the FHV RNA having a TYMV CP sequence at the 3'-end was efficiently encapsidated. When an eGFP gene was inserted into the B2 ORF of the FHV sequence, a fusion protein of B2-eGFP was produced as expected.

월경전후기증상의 유형과 월경전증상 관련식이요인 (Patterns of Perimenstrual Symptoms and Related Dietary Factors to Premenstrual Syndromes)

  • 김혜원
    • 여성건강간호학회지
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    • 제10권2호
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    • pp.162-170
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    • 2004
  • Purpose: The purpose of this study was to differentiate between women with three perimenstrual symptom severity patterns : premenstrual syndrome(PMS), premenstrual magnification(PMM), and low symptom(LS), and to explore the related dietary factors to premenstrual symptoms. Method: Women were asked to keep a diary record of perimenstrual symptoms and food intake for 50 days. Result: Symptom patterns were defined for 26 among 38 women ; Eight(21.1%) demonstrated a PMS pattern, three(7.9%) demonstrated a PMM pattern, and fifteen(39.5%) exhibited a LS pattern. There were significant differences in symptom scores during the premenstrual phase($x^2=19.30$, p=.000), menstrual phase($x^2=13.32$, p=.001), and post menstrual phase($x^2=9.93$, p=.007) for three groups. Protein, vit E, vit C, niacin, folic acid, and phosphorus in the premenstrual phase, and energy, and vit B6 in the menstrual period were significantly different between the three groups. Among dietary compositions, amino acids, lipids, fatty acids, saturated fatty acids, natrium, vit B6, niacin, and vit E were negatively related to PMS symptoms. Conclusion: Pattern of perimenstrual symptoms should be differentiated for individualized PMS management. As a more efficient diet assessment for PMS women, randomized nutritional analysis during the 3 phases of the menstrual cycle should be done and a replication study is necessary with a larger sample.

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Movement of Zucchini yellow mosaic vims Involved in Symptom Severity on Zucchini Squash

  • Park, Seung-Kook;Yoon, Ju-Yeon;Park, Sun-Hee;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • 제19권4호
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    • pp.217-220
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    • 2003
  • Zucchini squash (Cucurbita pepo cv. Black Beauty) plants infected with A strain of Zucchini yellow mosaic virus (ZYMV-A) isolated from a hollyhock plant showed systemically severe mosaic symptom, similar to previously established Cu strain of ZYMV. However, initial symptom of squash infected by ZYMV-A strain was generally more severe than those infected by ZYMV-Cu. Using leaf-detachment assay, examination of kinetics of accumulation of the coat protein (CP) in systemic loaves of squash plants showed that CPs of ZYMV-A appeared earlier than those of ZYMV-Cu. However, both ZYMV-A and ZYMV-Cu showed similar kinetics of CP accumulation 7 days post-inoculation. These results indicate that different rates and initial severity of systemic symptom development were due to differences in the rate of movement rather than vims replication.

식용견 문화의 변화와 진화론적 고찰 (Changes in Edible Culture of Dog Meat and Evolutionary Study)

  • 심순철;최현정
    • 한국조리학회지
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    • 제24권1호
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    • pp.122-129
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    • 2018
  • The purpose of this study is to understand the evolution of food culture by applying the evolutionary mechanism to the process of forming the dog meat culture. To do this, this study first examined mutation, selection, and replication as a evolutionary mechanism by biological genes and explain the evolutionary process of food culture by applying so-called 'mime' which is a virally-transmitted cultural symbol or social idea. A meme acts as a unit for carrying cultural ideas, practices, that can be transmitted from one mind to another through writing, speech, gestures, rituals, or other imitable phenomena with a mimicked theme. In addition, this study also intended to use in-depth interviews on how people have diverse cultural perspectives interpret and accept edible culture of dog meat. In Korea, which was a traditional farming society, dog meat which is easier to obtain compare to beef has been chosen as an important source of protein. And this choice has been repeatedly reproduced through generations. However, the current generation's awareness of the edible culture of dog meat has changed. The meme of pet culture has been selected and replicated, and this cultural evolution will eventually lead to the culling of dog meat.

분열형 효모에서 유전자 결실에 의해 알킬화제와 3-AMINOBENZAMIDE에 저항성을 나타내는 새로운 유전자의 특성 분석 (Characterization of a New Gene Resistant to Alkylating Agents and 3-Aminobenzamide When Knocked Out in Fission Yeast)

  • 박종군;차재영;황성진;박세근;김미영;백성민;최인순;이정섭
    • 생명과학회지
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    • 제12권2호
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    • pp.219-225
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    • 2002
  • 진핵세포의 염색체는 전사, 복제, 회복 등의 과정에서 관여하는 단백질의 기능으로 구조가 변하게 된다. 이때 관여하는 단백질은 DNA-단백질의 상호작용에 의해서 이루어지게 되는데, 이때 단백질의 일부분은 일정한 상동성이 존재하게 된다. 이러한 부분은 motif나 domain으로 구성되는데, 예를 들면, SAP domain등을 들 수 있다. S. pombe genomic DNA 데이터베이스를 검색하여 Arabidopsis PARP 과 KU70과 상동성을 보이는 새로운 유전자를 찾았다. 이를 SAPuvs (SAP UV Sensitive)라 명명하였으며, Ura4를 선별표지로 이용하여 S. pombe SAPuvs 유전자 결실세포를 구성하였다. SAPuvs 유전자 결실세포는 자외선 조사 실험에서 정상의 세포에 비해 현저하게 죽었다. 그러나, MMS 또는 MMS와 3AB의 처리 실험에서는 저항성을 보였다. 이러한 결과로 SAPuvs는 DNA 상해회복에서 염색사구조 형성에 연관되어 있음을 확인하였다.

Recombinant Adenoviral Vector Containing Tumor-Specific L-Plastin Promoter Fused to Cytosine Deaminase Gene as a Transcription Unit: Generation and Functional Test

  • Chung, In-Jae;Deisseroth, Albert-B.
    • Archives of Pharmacal Research
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    • 제27권6호
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    • pp.633-639
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    • 2004
  • The expression of therapeutic transgenes in recombinant adenoviral vectors is a major cause of toxicity in dividing cancer cells as well as non dividing normal cells. To solve the problem of toxicity to normal cells, we have reported on a recombinant adenoviral vector system (AdLP-) in which the expression of the transgene is directed by the tumor-specific L-plastin promoter (LP) (Chung et al., 1999). The object of this study was to generate a recombinant adenoviral vector system which would generate tumor cell specific expression of cytosine deaminase (CD) gene. We report the construction of a replication-incompetent adenoviral vector in which CD is driven by the L-plastin promoter (AdLPCD). Infection of 293 cells by AdLPCD generated the functional CD protein as measured by HPLC analysis for the conversion of 5-Fluorocy-tosine (5-FC) to 5-Fluorouracil (5-FU). HPLC analysis in conjunction with counting radioactivity for [6-$^3$H]-5FC and [6-$^3$H]-5FU demonstrated vector dose-dependent conversion of 5-FC to 5-FU in AdLPCD infected ovarian cancer cells. The results from present and previous studies(Peng et al., 2001; Akbulut et al., 2003) suggest that the use of the AdLPCD/5-FC system may be of value in the treatment of cancer including microscopic ovarian cancer in the peritoneal cavity.

Plasma Surface Modification of Patterned Polyurethane Acrylate (PUA) Film for Biomedical Applications

  • Yun, Young-Shik;Kang, Eun-Hye;Yun, In-Sik;Kim, Yong-Oock;Yeo, Jong-Souk
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2015년도 제49회 하계 정기학술대회 초록집
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    • pp.223.2-223.2
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    • 2015
  • Polyurethane acrylate (PUA) has been introduced to utilize as a mold material for sub-100 nm lithography as it provides advantages of stiffness for nanostructure formation, short curing time, flexibility for large area replication and transparency for relevant biomedical applications. Due to the ability to fabricate nanostructures on PUA, there have been many efforts to mimic extracellular matrix (ECM) using PUA especially in a field of tissue engineering. It has been demonstrated that PUA is useful for investigating the nanoscale-topographical effects on cell behavior in vitro such as cell attachment, spreading on a substrate, proliferation, and stem cell fate with various types of nanostructures. In this study, we have conducted surface modification of PUA films with micro/nanostructures on their surfaces using plasma treatment. In general, it is widely known that the plasma treated surface increases cell attachment as well as adsorption of ECM materials such as fibronectin, collagen and gelatin. Effect of plasma treatment on PUA especially with surface of micro/nanostructures needs to be understood further for its biomedical applications. We have evaluated the modified PUA film as a culture platform using adipose derived stem cells. Then, the behavior of stem cells and the level of adsorbed protein have been analyzed.

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