• 제목/요약/키워드: ${\beta}-galactosidase$ assay

검색결과 53건 처리시간 0.029초

Endocrine Disrupting Activity of Seven Phthalate Analogues in vitro

  • Ryu, Jae-Chun;Kim, Hyung-Tae;Kim, Youn-Jung;Jeon, Hee-Kyung
    • 한국환경성돌연변이발암원학회지
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    • 제22권4호
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    • pp.259-265
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    • 2002
  • Phthalate analogues are a plasticizer and solvent used in industry. Phthalates were reported to be a potential carcinogen classified in the category of suspected endocrine disruptors. Most common human exposure to these compounds may occur with contaminated food. They may migrate into food from plastic wrap or may enter food from general environmental contamination. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of phthalates that possibly threaten the public health. Concern about their use has been mounting. To screen and elucidate the endocrine disrupting activity and their mechanism of phthalate analogues, first of all, E-screen assay was performed in MCF7 human breast cancer cells with seven phthalate analogues. In this cell proliferation assay, only dibutyl phthalate (DBP) showed weak estrogenic activity. Also the yeast-based transcription assay to assess the interactions of DBP with the estrogen, androgen, and progesterone receptors was conducted. DBP in the concentration ranges from 10$^{-16}$ to 10$^{-11}$ M was active in the estrogen transcriptional assay, but it did not show the effect on $\beta$-galactosidase activity in the progesterone and androgen transcriptional assays. These data indicate that DBP shows estrogenic potential and can be classified as weak and/or suspected endocrine disrupting chemicals.

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Direct Analysis of the Transcription of Escherichia coli rnpB Gene Harbored in a Multicopy Plasmid during Bacterial Growth

  • Park, Jeong-Won;Jung, Young-Hwan;Park, Bo-Hyun;Jeoung, Yeon-Hee;Lee, Young-Hoon
    • BMB Reports
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    • 제29권3호
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    • pp.221-224
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    • 1996
  • To examine the growth-phase dependent control of Escherichia coli rnpB gene we used a combination of Northern analysis for RNA determination and Southern analysis for plasmid DNA determination. The relative amounts of metabolically unstable transcript derived from the internally deleted rnpB gene harbored on a multicopy plasmid as well as the relative plasmid contents were measured by Northern analysis and Southern analysis, respectively, of total nucleic acids from E coli cells containing the plasmid. The relative transcription activity of the rnB was represented by a ratio of the relative amount of the transcript to that of the plasmid DNA during bacterial growth. The rnpB transcription increased rapidly with time during exponential growth, but started to decrease before the transition period of an exponential growing cell culture into the stationary phase. Although the expression pattern was similar to the changes of ${\beta}-galactosidase$ activity expressed from the lysogenic strain carrying the chromosomal rnpB-lacZ fusion which were shown in a previous work, the present data appears to represent a more actual growth-phase control of the rnpB transcription than the previous data by the ${\beta}-galactosidase$ assay. In addition the present method described for a direct analysis of both RNA and plasmid DNA provides a rapid and efficient method that can applied to an examination of transcription control by using a multicopy plasmid.

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Ames 및 umu assay를 이용한 감궁탕의 안전성평가 (Evaluation of Safety with Gamgung-tang Using Ames and umu Assays)

  • 손윤희;김철호;남경수
    • 생명과학회지
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    • 제15권2호
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    • pp.215-219
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    • 2005
  • 감궁탕의 돌연변이원성을 유무를 알아보기 위해 Salmonella typhimurium TA 98 및 TA100을 이용한 돌연변이 원성 실험에서도 감궁탕은 어느 균에서도 돌연변이원성을 나타내지 않았으며, 이는 S-9 mixture 의해 감궁탕이 대사가 된 후에도 이와 유사한 경향을 나타내었다. 또한, SOS umu test의 경우에서도 $\beta-galactosidase$활성에는 별다른 영향을 미치지 않는 것으로 보아 감궁탕은 돌연변이원성을 일으키지 않는 것으로 판정되었으며 S-9 mixture처리에 의한 대사 후에도 이와 유사한 실험결과가 나타났다. 따라서 감궁탕은 그 자체 및 대사 후에도 DNA에 별다른 영향을 미치지 못하는 비교적 안전한 생약처방으로 여겨진다.

식용식물(食用植物)의 항변이원성(抗變異原性)에 관한 연구(硏究) (Assay of Antimutagenic Activities of Vegetable Plants)

  • 서정숙;이용욱;서난주;장일무
    • 생약학회지
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    • 제21권1호
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    • pp.88-91
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    • 1990
  • Potential antimutagenic activities of vegetable plants were investigated. 24 vegetables which are frequently consumed by Korean people were extracted with 70% ethyl alcohol to prepare the extract samples. Then those samples were added to the culture media containing mitomycin C($O.3\;{\mu}g/ml$) and the SOS-Chromotest was performed. Positive control, mitomycin C alone, showed about 330 units of ${\beta}$-galactosidase activities. Among 24 vegetable samples, Saxifraga oblougifolia Nakai (취나물, Saxifragaceae) and Platycodon grandiflorum A. De Candolle(도라지, Campanulaceae) showed 136 and 155 units, respectively when mitomycin C was treated. These results indicate that Saxifragae Herba and Platycodi Radix possess protecting action from mutagenic activity produced by mitomycin C.

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Enzyme immunoassay(EIA)에 의한 소의 혈중 progesterone 측정과 이의 응용에 관한 연구 I. 이항체(二項體)의 최적조건에 관한 연구 (Studies on enzyme immunoassay for determining progesterone of bovine plasma and its clinical application I. Optimizing double antibody for progesterone in enzyme immunoassay)

  • 강정부;신종욱;최상용
    • 대한수의학회지
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    • 제28권2호
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    • pp.321-325
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    • 1988
  • This experiment was carried out to determine the progesterone concentration of bovine plasma by liquid phase double antibody enzyme immunoassay. The optimum conditions of assay-system, double (first and second) antibody and carrier (normal rabbit serum) were investigated. The optimum dilution rate of first antibody, second antibody and normal rabbit serum was $10{\times}10^3$ to $15{\times}10^3$, 20 and $1{\times}10^3$ times, respectively.

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Ames, Rec 및 umu Assay를 이용한 황기의 안전성평가 (Evaluation of Safety with Astragali Radix : Ames, Rec and umu Assays)

  • 손윤희;남경수
    • 생약학회지
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    • 제34권1호통권132호
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    • pp.80-85
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    • 2003
  • Water extract from Astragali Radix (AR) was tested for the safety using Ames, Bacillus subtilis Rec, and umu gene expression mutagenicity tests. Mutagenic activity in any assays we tested was not found. In Ames test, Salmonella typhimurium TA98 and TA 100 were used to identify mutagenic property, and the number of histidine revertants was measured. In the Recassay, Bacillus subtilis ${H-17(Rec^+)\;and\;M-45(Rec^-)}$ strains were used to test DNA damage activity. In the SOS umu test, Salmonella typhimurium TA1535 containing plasmid pSK1002 was used as a test strain, and we monitored the levels of umu operon expression by measuring the ${\beta}-galactosidase$ activity. From the results, there was no DNA damage and mutagenicity of AR. Hepatotoxicity of AR to female ICR mice was also monitored by the measurements of s-GOT, s-GPT, LDH activities after oral feeding for 15 days. AR was not shown any significant changes of s-GOT, s-GPT and LDH activities in mice sera.

Transcriptional Regulation of Escherichia coli serC-aroA Operon : Further Support for cAMP-Dependent Expression

  • Sa, Jae-Hoon;Park, Soo-Sun;Lim, Chang-Jin
    • BMB Reports
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    • 제28권1호
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    • pp.21-26
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    • 1995
  • The Escherichia coli mixed-function serC-aroA operon encodes biosynthethic enzymes for unrelated pathways leading to the syntheses of serine and aromatic amino acids. It has been proposed that the operon is expressed in a cAMP-dependent manner. In this work experiments were performed to investigate the cAMP-dependent expression of the operon. Exogenous cAMP increased ${\beta}$-galactosidase synthesis in the $cya^+$ and cya strains harboring the serC-aroA-lac fusion plasmid. This enhancement was more dramatic in the $cya^-$ strain grown in a minimal medium. In a dot blot assay the serC-aroA mRNA content increased in a concentration-dependent pattern after the addition of exogenous cAMP. The activity of phosphoserine aminotransferase, encoded by the serC gene, apparently increased in E. coli cells after the addition of cAMP. All results obtained confirmed that the expression of the E. coli serC-aroA operon is positively regulated by cAMP at the level of transcription.

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변패미생물에 대한 녹차 물추출물의 항균 활성 분석 (Antimicrobial Activity of an Aqueous Extract of Green Tea against Food Putrefactive Microorganisms)

  • 신영희;오병태;최성길;허호진;이승철;조성환
    • 한국식품저장유통학회지
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    • 제16권3호
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    • pp.392-399
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    • 2009
  • 녹차는 생리활성의 촉진작용뿐아니라 항암, 항산화, 고혈압 억제효과, 체중조절 등 다방면의 약리효과를 보이는 것으로 보고되고 있다. 아울러, 녹차의 항균 및 항진균 작용에 관한 연구결과도 발표되고 있으나, 항균작용 기작에 접근한 기초 연구는 별로 많지 않다. 본 연구는 식품변패미생물에 대한 녹차추출물의 항균작용을 구명하는 기초자료를 획득하는데 그 목적이 있다. 녹차추출물의 항균력을 조사한 결과, $500{\mu}g/mL$ 이상의 농도에서 뚜렷한 항균활성을 보였고, 광역의 병원성 및 부패성 미생물들에 대하여 항균활성을 나타내었다. 또한, 녹차추출물의 항균물질은 넓은 범위의 온도($40{\sim}150^{\circ}C$) 및 pH($3{\sim}11$)에서 안정성을 나타냈다. 따라서 녹차추출물의 항균물질은 항균활성이 높고, 항균 대상 미생물의 분야가 광범위할 뿐아니라, 넓은 범위의 온도 및 pH에 안정하여 이상적인 천연 항균제로서의 개발가능성을 제시하였다. 아울러, 녹차추출물의 항균작용을 알기 위하여, 녹차추출물 처리가 미생물 세포막 기능에 미치는 영향을 조사한 결과, 녹차추출물의 주요 작용기작은 세포막 perturbation에 기인한 것으로 사료되었다.

어성초(魚腥草)의 항산화 효능 확인 및 모유두 세포의 5α-reductase 유전자 발현에 미치는 영향 (Investigation of Antioxidant Activity of Houttuyniae Herba and its Effect on 5α-reductase Gene Expression in Dermal Papilla Cells)

  • 조남준;이병권;이웅희;김기광;한효상
    • 동의생리병리학회지
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    • 제31권6호
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    • pp.356-361
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    • 2017
  • Houttuyniae Herba is widely used as a cosmetic for enhancing hair growth, and study on promoting mouse hair growth has also been reported. However, studies on the effects of the Houttuyniae Herba on dermal papilla (DP) cells, which play an important role in hair growth, are not well known. For this reason, we studied the effect of Houttuyniae Herba on DP cells. The strong antioxidant activity of Houttuyniae Herba was confirmed by ABTS assay. In the MTS assay, cell viability was reduced to 94.5% in DP cells by treatment of 2 mg/ml concentration of Houttuyniae Herb and cytotoxicity was not observed at 1 mg/ml concentration. The mRNA expression levels of Bone morphogenetic pretein (BMP6), fibroblast growth factor 7 (FGF7), FGF10, and ${\beta}$-galactosidase genes, which are involved in hair growth cycle and hair loss induction, were measured by quantitative RT-PCR after Houttuyniae Herbtreatment. Houttuyniae Herb did not significantly affect mRNA expression of BMP6, FGF7, FGF10, and ${\beta}$-catenin, which are important factors for regulating the hair cycle, including type 1 $5{\alpha}$-reductase. However, mRNA expression of type 2 $5{\alpha}$-reductase, the major cause of male hair loss, was significantly reduced to 56.1% by treatment of Houttuyniae Herbtreatment. Taken together, these results suggest that the Houttuyniae Herbtreatment can help to treat lair loss through removing free radicals and suppression of the expression level of type 2 $5{\alpha}$-reductase in DP cells.

재조합 Agrobacterium tumefaciens NTL4 균주를 이용한 quorum Sensing Autoinducer 검색에 용매와 계면활성제가 미치는 영향 (Effect of Solvents and Surfactants on the Whole-cell Bioassay for Screening Quorum Sensing Autoinducers Using the Recombinant Agrobacterium tumefaciens NTL4 Strain)

  • 고경표;김연희;김정선;박성훈
    • 한국해양바이오학회지
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    • 제1권4호
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    • pp.260-267
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    • 2006
  • 재조합 Agrobacterium tumefaciens균주를 사용한 Liquid Culture Assay는 quorum sensing autoinducer를 검색하는 생물학적 분석 방법으로 개발되었다. 그러나 이 시스템에 사용되는 해양천연물 시료들이 일반적으로 낮은 수용액에 대한 용해도를 갖기 때문에 활성검색에 걸림돌이 되고 있다. 시료의 용해도는 유기용매 혹은 계면활성제의 첨가로 증가될 수 있으나, 유기용매나 계면활성제 자체가 검색결과의 정확한 해석을 방해할 수 있으므로 적절한 조건의 확립이 매우 중요하다. Methanol, ethanol, 1-propanol, DMSO와 DMF를 0~10% 농도 범위에서 재조합 A. tumefaciens균주에 대한 세포 성장에 미치는 영향과 ${\beta}$-galactosidase activity에 미치는 영향을 살펴본 결과 methanol 2%이하의 농도가 가장 적합하였으며, 계면활성제의 경우, Tween 20, Tween 80보다는 Triton X-100이 약 0.05% 농도에서 세포내로의 활성물질의 전달효율을 높이는데 효과적이었다.

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