• 제목/요약/키워드: high-level expression

검색결과 1,212건 처리시간 0.028초

합성유전자를 이용한 식물단백질의 향상 (Plant Protein Improvement by Synthetic Gene)

  • 김태금;양문식
    • KSBB Journal
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    • 제7권3호
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    • pp.155-160
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    • 1992
  • 식량으로 쓰이는 식물 단백질은 공통적으로 Isoleucine, Lysine, Methionine, Threonine, Tryptoplan등 5가지 필수아미노산이 결핍되어있다. 본 연구에서는 이러한 필수아미노산을 다량 함유한 단백질을 발현시킬 수 있는 합성유전자를 fekaqo에서 높은 수준으로 발현시키고자 강한 식물 promoter로 알려진 CaMV 35S, CaMV duplicate 35S promoters를 사용하였다. 형질 전환 및 재분화된 식물을 분석한 결과 본 합성 유전자가 식물 nuclear genome 안으로 도입을 안정하여 잘되었고 mRNA수준까지는 유의적인 증가를 보였으나 단백질 수준에서는 유의적 수준의 증가를 관찰할 수 없었다.

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철 과잉투여가 흰쥐의 Hepatic Fibrogenesis와 Collagen 및 TGF-$\beta$I 유전자 발현에 미치는 영향 (Influence of Collagen and TGF-$\beta$I Gene Expression and Hepatic Fibrogenesis by Iron Overload in Rat)

  • 양영목;박종환;이현영;정연희;김해영
    • 한국식품영양과학회지
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    • 제30권2호
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    • pp.307-313
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    • 2001
  • Iron excess is known to affect long-term iron accumulation and tissue change such as fibrosis in liver. To determine the changes of expression level of genes associated with fibrosis by short-term iron exposure, we measured liver mRNA levels by reverse transcription polymerase chain reaction (RT-PCR) in rats fed dietary carbonyl iron (3%, wt/wt) for 9 weeks. The results showed that the expression of the collagen (I, III) and transforming growth factor (TGF)-$\beta$I mRNAs was enhanced in high-dose iron treated rat, compared to normal-dose iron treated rat. An electron microscopy study revealed that excess iron caused increase of collagen fibrils in liver. The cell shapes and compositions of hepatocytes and extracellular matrix(ECM) in liver were changed by the iron-treatment. Also, necrosised hepatocytes were broadly seen in ECM. Taken together, we suggest that iron overload affects changes of collagen and TGF-$\beta$I gene expression and these changes are associated with liver fibrogenesis.

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Molecular Effects of Genistein on Proliferation and Apoptosis of MCF-7 Cell Line

  • Shin, Hye-Jin;Oh, Young-Jin;Hwang, Seung-Yong;Yoo, Young-Sook
    • Molecular & Cellular Toxicology
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    • 제2권1호
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    • pp.15-20
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    • 2006
  • Genistein is a potent, plant-derived isoflavone that displays estrogenic activity at low concentrations but inhibits proliferation at high amounts. However, the molecular mechanism of genistein is not completely understood. In the present study, the biphasic effects (estrogenic and antiestrogenic activity) of genistein on the growth of MCF-7 cells were identified. Genistein within a low range of concentration, $1-10\;{\mu}M$, stimulated proliferation, while $50-100\;{\mu}M$ caused apoptotic cell death. Additionally, genistein at a low concentration induced estrogen receptor (ER)-mediated gene expression and ER phosphorylation. When pre-treated with PD98059, an MEK inhibitor, ER-mediated gene expression and ER phosphorylation by genistein were noticeably increased. However, the increased gene expression and phosphorylation did not enhance cell proliferation. Moreover, it was observed that ER-mediated signaling performs an important role in the MAPK pathway. The proliferation and apoptosis in genistein-treated MCF-7 cells were partially dependent on the Bcl-2 level. The addition of IC1 182, 780, an estrogen receptor antagonist, inhibited Bcl-2 expression induced by genistein. This study suggests that there is a close relationship between Bcl-2 and the ER signaling pathways in MCF-7 cells.

The Function of Multiple Pribnow Box on the Aerobic-Anaerobic Switch Control of aeg-46.5 Gene Expression

  • 강인오;정연주;최무현
    • Bulletin of the Korean Chemical Society
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    • 제22권8호
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    • pp.903-908
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    • 2001
  • The gene aeg-46.5, which is expressed under anaerobic condition, has putative triple -10 regions and four transcription start sites. The mRNA transcription level and its start point change depending on the aerobic/anaerobic growth condition. RNA polymerase and its regulatory proteins must choose which of three -10 region to use. The putative triple 10 region was mutated to make only one of them function with consensus -10 region sequence (TATAAT) and the other two as non-functional region. The results show that the second and third -10 regions are used for the aerobic/anaerobic expression. The third -10 region is responsible for the high aerobic to anaerobic switch ratio. This suggests that only the last two of the putative triple -10 region have functions on aeg-46.5 gene expression switch control. The phenotype of the mutated promoter was tested in the wild type cell and narL - cell. The results indicate that the control by NarL is independent from the selection of -10 region. The expression patterns on multi-copy plasmids and on single-copy chromosome were compared. These results show that the aerobic/anaerobic switch control of aeg-46.5 is through the choice of -10 region. The mechanism of choosing different -10 region remains to be seen.

Daidzein이 benzo(k)fluoranthene에 의한 사람 유방암 세포 MCF-7의 CYP1A1 유전자 발현 조절에 미치는 영향 (Effects of Daidzein on benzo(k)fluoranthene Regulated CYP1A1 Gene Expression in MCF-7 Human Breast Cancer Cells)

  • 양소연;신윤용
    • 한국환경성돌연변이발암원학회지
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    • 제24권4호
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    • pp.180-188
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    • 2004
  • CYP1A1 is known to be inducible by xenobiotic compouds such as polyciclic aromatic hydrocarbons(PAHs) and 2,3,7,8-tetrachloro-dibenzo-p-dioxin(TCDD). These chemicals have been identified worldwide and can have a significant impact on the human health and well being of human and wildlife. Given these issues, the detection and quantification of these chemicals in biological, environmental and food samples is important. We investigated the effect of dietaty flavonoid, such as CYP1A1 promoter activity, 7-ethoxyresorufin-O-deethylase(EROD) activity and CYP1A1 mRNA expression induced by benzo(k)fluoranthene(B(k)F) in MCF-7 cells. Based on the three criteria of frequency of occurrence in the environment, toxicity and potential exposure to humans, B(k)F is one of the top-listed PAHs. We found that B(k)F significantly up-regulates the level of CYP1A1 promoter activity, EROD and CYP1A1 mRNA. when cells were treated with daidzein inhibited the B(k)-induced CYP1A1 prompter activity and mRNA level at high concentration. But daidzein exhibited stimulatory effects B(k)F-induced CYP1A1 promoter activity and mRNA level at low concentration. Overall, results from these studies demonstrate flavonoids might interfere the action of B(k) with AhR system to stimulate CYP1A1 gene expression.

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Morin이 benzo(k)fluoranthene에 의한 유방암 세포 MCF-7의 CYP1A1 유전자 발현 조절에 미치는 영향 (Effects of Morin on benzo(k)fluoranthene Regulated CYP1A1 Gene Expression in MCF-7 Human Breast Cancer Cells)

  • 양소연;김여운;신윤용
    • 한국환경성돌연변이발암원학회지
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    • 제24권4호
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    • pp.189-197
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    • 2004
  • We investigated the effect of dietaty flavonoid, such as CYP1A1 promoter activity, 7-ethoxyresorufin-O-deethylase(EROD) activity and CYP1A1 mRNA expression induced by benzo(k)fluoranthene(B(k)F) in MCF-7 cells. Based on the three criteria of frequency of occurrence in the environment, toxicity and potential exposure to humans, B(k)F is one of the top-listed PAHs. We found that B(k)F significantly up-regulates the level of CYP1A1 promoter activity, EROD and CYP1A1 mRNA. When cells were treated with morin alonem, it was not changed that EROD and CYP1A1 mRNA, compared to that of control. However, morin inhibited the B(k)-induced CYP1A1 prompter activity and mRNA level at high concentration. But morin exhibited stimulatory effects B(k)F-induced CYP1A1 promoter activity and mRNA level at low concentration. Overall, results from these studies demonstrate morin might interfere the action of B(k) with AhR system to stimulate CYP1A1 gene expression. CYP1A1 is known to be inducible by xenobiotic compouds such as polyciclic aromatic hydrocarbons(PAHs) and 2,3,7,8-tetrachloro-dibenzo-p-dioxin(TCDD). These chemicals have been identified worldwide and can have a significant impact on the human health and well being of human and wildlife. Given these issues, the detection and quantification of these chemicals in biological, environmental and food samples is important.

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대사증후군이 있는 당뇨병 환자의 지각된 스트레스에 따른 정신건강과 자가당뇨관리 (Mental Health and Self-care Activities according to Perceived Stress Level in Type 2 Diabetic Patients with Metabolic Syndrome)

  • 김춘자
    • 성인간호학회지
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    • 제22권1호
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    • pp.51-59
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    • 2010
  • Purpose: This study was to identify the mental health and self-care activities according to perceived stress level in type 2 diabetic patients with metabolic syndrome. Methods: The descriptive survey design was conducted using a convenient sample. One hundred and fifty-two diabetic participants with metabolic syndrome were recruited in a university hospital. The data were analyzed using descriptive statistics, Students' t-test, and ANOVA with SPSS/WIN 12.0. Results: The high, moderate, and low stress groups according to perceived stress score consisted of 30.9%, 38.2%, and 30.9%, respectively. Differences of mental health and total self-care activities among the three groups were statistically significant. Depression (F=46.73, p<.001), trait anger (F=5.75, p<.01), and anger expression-in (F=4.60, p<.01) of participants in the high stress group were higher than other groups, whereas the total of self-care activities (F=2.85, p<.05) and exercise (F=3.49, p<.05) were lower than other groups. Conclusion: This study concluded that type 2 diabetic patients with metabolic syndrome with a high stress level were low in self-care activities along with elevated depression and anger.

고지방 식이를 섭취시킨 흰쥐에서 고추장의 항비만효과 (Anti-obesity Effects of Kochujang in Rats Fed on a High-fat Diet)

  • 주종재
    • Journal of Nutrition and Health
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    • 제33권8호
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    • pp.787-793
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    • 2000
  • The purpose of the present study was to investigate effects of kochujang and red pepper on energy intake, body fat content and energy expenditure in rats fed on high-fat(30%) diet. Kochujang and red pepper power were added in the high-fat diet, adjusting the level to 95 and 22g/kg diet, respectively, The level of red pepper addition was corresponding was corresponding to the level of the content of red pepper powder in the kichujang-added diet. Kochujang induced a 30% reduction in body fat gain which was associated with a significant increase in energy expenditure. However, red pepper reduced body fat gain by only 15%. Furthermore, energy expenditure was not affected by red pepper. Metabolizable energy intake, apparent digestibility and body protein gain were not affected by either kochujang or red pepper. It has been known that capsaicin, a pungent component of red pepper, enhances activity of brown adipose tissue through increasing protein content. In the present study, in addition of protein content, DNA content of interscapular brown adipose tissue was also increased by kochujang. Therefore, it appeared that the anti-obesity effects of kochujang was greater than those of red pepper, indicating more than red pepper was involved in the expression of the anti-obesity effects of kochujang(Korean J Nutrition 33(8) : 787-793, 2000)

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The Differential Staging of Murine Thymic Lymphoma Cell Lines, Scid.adh, R1.1 and EL-4

  • Chae, Jong Seok;Kim, Hae-jung;Park, Weon Seo;Bae, Youngmee;Jung, Kyeong Cheon
    • IMMUNE NETWORK
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    • 제2권4호
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    • pp.217-222
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    • 2002
  • Background: Scid.adh is a recently developed murine thymic lymphoma cell line, which has been used as in vitro model for the study of double negative stage III thymocytes. In this study, we compared the expression profile of a number of genes and proteins, which are tightly related to T cell development and apoptosis, in thymic lymphoma cell lines, R1.1, EL-4, and Scid.adh for the developmental staging. Methods: We examined the expression of development marker genes and proteins in three lymphoma cell lines by flow cytometry and RT-PCR. In addition, the expression of apoptosis-related molecules including bcl-2, bax and Fas was also investigated. Results: As previously reported, Scid.adh cell line expressed CD8 and CD25 but not TCR ${\alpha}$ chain, while R1.1 cells expressed TCR ${\alpha}$ chain and both CD4 and CD8 transcripts. These suggest that R1.1 might be in double positive stage, and low level of CD44 expression and the absence of CD25 support this suggestion. In contrast, EL-4 cells showed high level of TCR ${\alpha}$ chain transcript, and low-level of CD4 expression, suggesting that EL-4 is in more mature stage than R1.1. Further, this suggestion was supported by the lack of mT-20 in EL-4 cells, which is expressed in the immature thymocytes, and Scid.adh and R1.1 cell lines, but not in the terminally differentiated thymocytes and peripheral T cells. Among the apoptosis-related gene, transcripts of bcl-2 gene were detected in both R1.1 and EL-4 but not in Scid.adh cells, while bax was expressed in all cell lines. Fas expression was the highest in EL-4 cells and low in Scid.adh cell line. Conclusion: R1.1 cell may represent double positive stage, and EL-4 is more differentiated cell line. In addition, Scid.adh and EL-4 cell lines are suspected to be useful for the study of function of bcl-2 family and Fas during the thymocyte development, respectively.

MC3T3-E1 세포의 골기질 단백질 발현에 대한 혈소판유래성장인자-BB의 효과 (The Effects of Platelet- Derived Growth Factor-BB on the Expression of Bone Matrix Protein in the MC3T3-E1 Cells)

  • 김묘선;이재목;서조영
    • Journal of Periodontal and Implant Science
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    • 제30권2호
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    • pp.347-360
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    • 2000
  • Bone remodeling results from the combined process of bone resorption and new bone formation which is regulated in part by some of the polypeptide growth factors such as platelet derived growth factor(PDGF), which has been known to be an important local regulator of bone cell activity and participate in normal bone remodeling. This process includes strictly regulated gene expression of several bone matrix proteins such as type I collagen and osteopontin, a 44 kDa phosphorylated glycoprotein, which has important roles in bone formation. The purpose of this study is to evaluate the effecs of PDGF-BB on the mRNA expression of bone matrix protein, type I collagen and osteopontin, in MC3T3- E1 cell culture. Cells were seeded at $5{\times}10^5$ cells in 10 ml of minimum essential medium alpha(${\alpha}-MEM$) containig 10% fetal bovine serum, 10 mM beta glycerophosphate. 0.1, 1, 10 ng/ml PDGF-BB were added to the cells for the day 3, 7, 14, 21, 28 and cultured for 24 hours. Type I collagen cDNA, Hf677, and osteopontin cDNA were used as probes for northern blot analysis. Total cellular RNA was purified at indicated day and northern blot analysis was performed. The results were as follows : Type I collagen mRNA expressions were higher at the day 3 and 7, and lower in the day 14, 21 in the control groups. In the experimental groups, mRNA expressions were increased when 0.1 ng/ml PDGF-BB were added on the day 3, 7, 21, and decreased in dose-dependent manner on the day 14, decreased at all added dose on the day 28. Osteopontin mRNA expressions were highest in the day 21 groups and lowest in the day 14 groups in the control groups. Interesting results were shown in the day 14 and 21 groups. We found that osteopontin mRNA level was increased in dose dependent manner in the day 14 groups, and decreased dose dependent manner in the day 21 groups. In conclusion, PDGF-BB may have various control effects on type I mRNA expression in the growth and differentiation process of MC3T3-E1 cells and may have contrary regulatory effects on osteopontin mRNA expression. For examples, when the baseline level of osteopontin mRNA was low, as in the day 14, PDGF-BB up-regulated osteopontin mRNA expression in dose dependent manner, and when the baseline level was high as in the day 21, PDGF-BB down-regulated dose dependent manner. Thus, it may be useful for clinical application in periodontal regeneration procedure if further study were performed.

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