• 한국양식학회지
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• 제13권1호
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• pp.29-37
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• 2000

In previously study we isolated several fish matrix attachment regions (MARs) capable of replicating the plasmid by itself. In this study we construct a fish expression vector pBaEGFP(+) containing mud loach ${\beta}$-actin promoter EGFP as reporter gene and SV40 signal. To analyze the effects of the fish expression vector respectively. The fish ARS containing constructs pBaEGFP(+)-ARSs were transfected cells with pBaEGFP(+)-ARS101 and pBaEGFP(+)-ARS223 reduced 10 days to 25 days and then was constant to 30 days after transfection while that of the control vector without ARS element was basal level. The intensity of both constructs showed about 30fold of the intensity compared with the control vector on 30days after transfection individually .E. coli back-transformation analysis shows that pBaEGFP(+)-ARS223 and pBaEGFP(+)-ARS905 maintain in episomal state at least 30 days after transfection. The result indicates that both may be able to replicate the vector in BF-2 cell. Therefore the matrix-attached ARSs enhancing expression of the reporter gene might be useful as a component o the expression vector for transgenic studies.

• 복식
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• 제56권8호
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• pp.148-159
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• 2006

The objective of this study is to consider the uniform as visual communication, and analyze the expression of symbol shown on the uniform design by the pattern by applying the visual symbol in the communication and operation of the meaning. Also, it is to construct the systematic modeling method of the expression of design symbol by the pattern of uniform in th e various fields. This study is to analyze the symbolic expression method of the uniform design by the pattern by suggesting the symbolic expression method as a frame through the visual symbolic approach of the uniform design and the analysis of semantic symbol according to that. As a result, there arose simplification and emphasis and a method of symbolic expression with the national flag, a conceptual factor characteristically in the sports uniform design. The characteristics of symbolic expression of uniform design were mainly used in the symbolic expression with the customary code and symbol. Also, the uniform in the enterprises were widely used in the expression of uniform with the suggestion and code by the color. As for the characteristics of symbolic expression shown on the event uniform design, the method of symbolic expression by the pattern of uniform was grasped differently asa result of symbolic expression through the image. The method of symbolic expression suggested on this study could be used on a basis of uniform design in the future. Accordingly, this study has a meaning of suggesting the methodology of uniform design that would be a parameter.

• Reproductive and Developmental Biology
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• 제32권2호
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• pp.135-140
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• 2008

Recent studies on nuclear transfer and induced pluripotent stem cells have demonstrated that differentiated somatic cells can be returned to the undifferentiated state by reversing their developmental process. These epigenetically reprogrammed somatic cells may again be differentiated into various cell types, and used for cell replacement therapies through autologous transplantation to treat many degenerative diseases. To date, however, reprogramming of somatic cells into undifferentiated cells has been extremely inefficient. Hence, reliable markers to identify the event of reprogramming would assist effective selection of reprogrammed cells. In this study, a transgene construct encoding enhanced green fluorescent protein (EGFP) under the regulation of human Oct4 promoter was developed as a reporter for the reprogramming of somatic cells. Microinjection of the transgene construct into pronuclei of fertilized mouse eggs resulted in the emission of green fluorescence, suggesting that the undifferentiated cytoplasmic environment provided by fertilized eggs induces the expression of EGFP. Next, the transgene construct was introduced into human embryonic fibroblasts, and the nuclei from these cells were transferred into enucleated porcine oocytes. Along with their in vitro development, nuclear transfer embryos emitted green fluorescence, suggesting the reprogramming of donor nuclei in nuclear transfer embryos. The results of the present study demonstrate that expression of the transgene under the regulation of human Oct4 promoter coincides with epigenetic reprogramming, and may be used as a convenient marker that non-invasively reflects reprogramming of somatic cells.

• 원예과학기술지
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• 제31권1호
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• pp.95-102
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• 2013

이원적 전사유도 시스템(binary trans-activation system)은 도입유전자의 발현을 조절하는 기작(mechanism) 중에 하나로, 목적 유전자의 발현이 전사활성 인자를 가지고 있는 식물체와의 교배를 통해서만 발현되는 시스템이다. 본 연구에서는 이원적 전사유도 시스템을 원예 작물의 우수한 유전자원 및 신품종 보호 방법으로 이용하고자, 배추에서 이 시스템의 기능을 검정하였다. 배추작물에서 이원적 전사유도 시스템의 이용가능성을 검정하기 위하여 activator construct(35SLhGBart)와 reporter construct(pOpGUS1300)를 작성하였고 공동형질전환방법으로 배추에 형질전환하였다. 두 종류의 카세트가 도입된 형질전환체는 항생제를 이용하여 선발하였으며, 재분화된 신초의 GUS 유전자 발현으로 이 시스템의 활성을 확인하였다. 또한 이 시스템을 조직 특이적으로 유도하기 위하여 애기장대의 자성 배우체 특이적 프로모터를 이용하여 activator construct(795LhGBart)를 작성하여 애기장대에 형질전환 하였다. 공동형질전환된 애기장대는 자성 배우체에서 조직 특이적인 발현을 나타냈다. 이러한 결과는 이원적 전사유도 시스템이 목적유전자의 발현을 배추의 $F_1$ 종자에서 선택적으로 유도하는 방법으로써 우수한 유전자원 및 신품종 보호에 이용될 수 있다는 것을 보여주는 것이라고 생각된다.

• International Journal of Industrial Entomology and Biomaterials
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• 제4권1호
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• pp.51-56
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• 2002

A recombinant transfer vector pBacSCF was constructed by inserting huamn stem cell factor (hSCF) cDNA into plasmid pBacPAK8. BmN cells were co-transfected with modified Bombyx mori, nuclear polyhedrosis virus (BmBacPAK) DNA and the recmbinant transfer vector to construct a recombinant baculovirus containing hSCE gene. DNA dot blotting and RNA dot blotting demonstrated that the hSCE gene was contained in the recombinant virus and transcribed. The recombinant baculovirus was infectious to BmN cells and to silkworm. SDS-PAGE analysis showed a specific band of expressed product in the extract of infected cells and in the heamolymph of infected larvae. Bioactivity of the recombinant hSCE was determined with W-1 cell line and MTT colorimetric method in synergy with interlukin-3 (IL-3). These results revealed that the hSCF gene was over-expressed in cultured cells and lavae of silkworm.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 1998년도 제4차 학술발표대회 및 정기총회
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• pp.59-60
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• 1998

A DNA construct containing rat T $\alpha$ 1 $\alpha$ -tuulin gene 5'-flanking sequence and GFP reporer gene was microinjected into 1-cell loach embryos. Neuron-specific FGP expression was observed in developing loach embryos and early stage fry. The results demonstrated that rat T $\alpha$ 1 $\alpha$ -tubulin gene promoter may be sufficient to specify gene expression to neurons in loach embryos. Thus, the use of GFP reporter controlled by T $\alpha$ 1 $\alpha$ -tubulin gene promoter may facilitate visualization of the dynamic processes of neural tissue development.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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• pp.189-189
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• 2001

For a continuous expression of human cytochrome p450 3A5 (CYP3A5) and NADPH-cytochrome P450 reductase (CYPR) proteins, bicistronic construct (CYP3A5BC-LNCX2) was made using internal ribosome entry site (IRES). As for mammalian cell expression, we used pLNCX2 retroviral vector; and using calcium phosphate, plasmid transfer was achieved in 293GPG cell and transduced in Chinese hamster ovary (CHO) cell.(omitted)

• BMB Reports
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• 제41권8호
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• pp.568-574
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• 2008

Antisense RNA molecules are powerful tools for controlling the expression of specific genes but their use in prokaryotes has been limited by their unpredictable antisense effectiveness. Moreover, appreciation of the molecular mechanisms associated with silencing in bacteria is still restricted. Here we report our attempts to define an effective antisense strategy in E. coli, and to dissect the observed silencing process. Antisense constructs complementary to different regions of lacZ were investigated, and silencing was observed exclusively upon expression of antisense RNA hybridising the 5'UTR of lac messenger. The level of lacZ mRNA was reduced upon expression of this antisense construct, and the silencing competence was found to be closely associated with its stability. These observations may help in the design of antisense molecules directed against prokaryotic genes.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.140-140
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• 1998

The aim of this study was to find out the effect of hypoxic condition on the regulation of cyplal gene expression. pcyplal-Luc construct was cloned and transfected into Hepa I cells. When Hepa-I cells containing pcyplal-Luc were treated by DFO (desferrioxamine) which is iron-chelating agent, the stimulatory effect of luciferase by TCDD was decreased. This inhibitory effect of desferrioxamine on the luciferase activity was dose dependent and abolished by concomitant treatment with N$\^$G/-nitro-ι-arginine. And when cobalt chloride which is known as a hypoxia inducing chemical was administrated, the stimulatory effect of luciferase by TCDD was also decreased. This inhibitory effect of cobalt chloride on the luciferase activity was dose dependent and abolished by concomitant treatment with N$\^$G/-nitro-ι-arginine. These data showed that hypoxic condition down regulates cyplal gene expression and this might be through nitric oxide action.

• BMB Reports
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• 제36권5호
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• pp.493-498
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• 2003

The scFv antibody towards the Burkholderia pseudomallei exotoxin was previously constructed by phage display and exhibited good specificity towards the exotoxin. We report here the optimization of the scFv expression in an E. coli expression system. Four different E. coli strains (ER2537, TG1, HB2151, and XL1-Blue) were examined for optimal expression of the scFv protein. Two types of carbon source (i.e. 0.2% glucose and 0.2% glycerol) were also tested for their ability to induce the scFv expression. Cells that carried the scFv construct were grown at $30^{\circ}C$ and induced with 0.05 mM IPTG. The expression was then monitored by SDS-PAGE, Western blotting, and indirect ELISA. The Western blot profile showed different levels of the scFv expression among the host strains; XL1-Blue exhibited the highest level of the scFv protein expression. Glycerol at a concentration of 0.2% (v/v) significantly increased the scFv protein expression level when compared to 0.2% (w/v) glucose. Further optimization demonstrated that the scFv protein expression in XL1-Blue was the most optimal with a glycerol concentration as low as 0.05%. However, by indirect ELISA, only the scFv protein that was expressed in 0.2% (v/v) glycerol exhibited high specificity towards the Burkholderia pseudomallei exotoxin.