• 제목/요약/키워드: aP2 promoter

검색결과 704건 처리시간 0.028초

Resveratrol에 의한 cysteine-rich angiongenic inducer 61 (CYR61) 유전자의 과대발현 조절 (Resveratrol Up-regulates Cysteine-rich Angiogenic Inducer 61 (CYR61) in Human Colorectal Cancer Cells)

  • 곽은희;김종식
    • 생명과학회지
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    • 제23권2호
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    • pp.207-212
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    • 2013
  • 파이토케미칼 resveratrol은 항산화, 항염증, 항암등을 포함하는 다양한 생리활성을 가지고 있는 것으로 알려져 있다. 본 연구에서는 resveratrol이 CCN family 중의 하나인 cysteine-rich angiogenic inducer 61 (CYR61) 유전자의 발현을 유도할 수 있는지 연구하였다. 결과에 의하면 resveratrol은 3개의 다른 인간 대장암 세포주에서 CYR61 단백질의 발현을 유도하였을 뿐만 아니라, HCT116세포주에서는 처리한 resveratrol 농도와 시간 의존적으로 CYR61 단백질의 발현을 유도하였다. 이러한 CYR61 단백질의 발현이 resveratrol의 어떤 생리활성과 관련이 있는지 확인하기 위하여 몇 종류의 NSAIDs와 항산화제를 처리하여 CYR61 단백질의 발현을 확인하였으나, 오직 resveratrol의 처리에 의해서만 CYR61 단백질의 발현이 유도되었다. 또한, CYR61의 발현은 암 억제유전자인 p53과는 관련이 없는 것으로 판단되었다. Promoter assay를 통하여 프로모터 -732 ~ +54 사이에 조절부위가 있음을 확인하였고, 파이토케미칼 Indole-3-carbinol이나 6-gingerol에 의해서도 CYR61의 발현이 유도되지 않음을 확인하였다. 이러한 연구결과는 resveratrol에 의한 CYR61 유전자의 발현은 resveratrol특이적이며, 이러한 연구결과는 resveratrol만의 특이한 생리활성을 이해하는데 도움을 줄 것으로 기대된다.

Production of Monoclonal Antibody about Specific Key Enzyme of Hyoscyamine $6{\beta}-Hydroxylase$ (H6H) in Scopolia parviflora

  • Kang, Young-Min;Jung, Hee-Young;Kang, Seung-Mi;Jin, Byung-Rae;Lee, Sang-Chul;Lee, Byung-Hyun;Choi, Myung-Suk
    • 한국약용작물학회지
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    • 제12권2호
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    • pp.135-140
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    • 2004
  • Total RNAs were isolated from cultured roots of Scopolia parviflora, $poly(A)^+$ RNA was obtained through the mRNA purification, cDNA library of Hnh6h was constructed. Recombinant baculoviruses in Spodoptera frugiperda (Sf) cells were constructed by use of the transfer vector pBacPAK, which has the AcNPV sequence under the polyhedrin promoter. The expression vector carrying Hnh6h gene was transferred to S. parviflora and obtained transgenic hairy root lines. Our results confirmed the over expression of the H6H protein was used by anti-pBacPAK about cDNAs of S. parviflora. This study will served for production of tropane alkaloids by metabolic engineering.

Characteristics of fermented milk containing Bifidobacterium growth promoter (BE0623) and dietary fiber

  • Cho, Young Hoon;Sim, Jae Young;Nam, Myoung Soo
    • 농업과학연구
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    • 제48권2호
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    • pp.209-218
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    • 2021
  • This study was carried out to investigate the effects of Bifidobacteria growth promoter BE0623 and a dietary fiber supplement, which included Bifidobacterium lactis BB12, Lactobacillus acidophilus, Streptococcus thermophilus, and Bifidobacterium lactis. In fermented milk containing BE0623, the viable cell count of Bifidobacteria significantly increased by about 45 to 75 times compared to the control, and the titratable acidity increased, whereas the pH decreased. All fractions obtained by isolating BE0623 had Bifidobacteria growth effect. Acacia dietary fiber is a pale yellow powder. It has a viscosity of 60 to 100 cPs and a pH between 4.1 and 5.0. Its general components are less than 10% moisture, more than 90% dietary fiber, and less than 4% ash. The optimal addition ratio of Bifidobacteria growth promoting material was determined to be 0.05%. The general components of the manufactured fermented milk were carbohydrate 17.85%, protein 3.63%, fat 3.00%, and dietary fiber 2.95%. During storage of the fermented milk for 24 days, its titratable acidity, viscosity, and sugar content all met the criteria. In addition, the viable cell counts of Bifidobacteria and lactic acid bacteria in the fermented milk were 1.7 × 108 CFU·mL-1 and 1.5 × 107 CFU·mL-1, respectively, and Escherichia coli was negative. There was no significant difference between the control group and the treatment group in the sensory evaluation of sweet, sour, weight, and flavor, and the preference for the treatment group was excellent. The acceptability of the fermented milk of the treated group according to the storage period was excellent in terms of color, flavor, and appearance.

Cytochrome P-450 3A4 Proximal Promoter Activity by Histone Deacetylase Inhibitor in Hepg2 Cells

  • Kim, Ja-Young;Ahn, Mee-Ryung;Sheen, Yhun-Yhong
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2003년도 추계학술대회
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    • pp.166-166
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    • 2003
  • Cytochrome P-450 3A4 (CYP3A4) is major enzyme in human liver, the role of this is detoxification and metabolizing more than 50% clinical drugs in use. The transcription of CYP3A4 is regulated by the Pregnenolone X receptor (PXR),of which human form is Steroid and Xenobiotics receptor (SXR).(omitted)

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E. coli에서 Pseudoalteromonas carageenovora 유래 Arylsulfatase의 구성적 발현과 Agarose 제조에의 응용 (Constitutive Expression of Arylsulfatase from Pseudoalteromonas carageenovora in E. coli and Its Application to Preparation of Agarose)

  • 김미진;장연화;성문희;김연희;남수완
    • 한국미생물·생명공학회지
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    • 제35권1호
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    • pp.11-16
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    • 2007
  • Pseudoalteromonas carrageenovora 유래의 arylsulfatase 유전자는 PCR로 증폭한 후 Geobacillus toebii의 D-amino acid aminotransferase(D-ATT) 유전자 유래의 구성적 발현 promoter를 함유하는 pHCE-IA vector로 subcloning 하였다. 4-Methylumbelliferyl sulfate가 포함된 LB 평판배지 상에서 자란 형질전환체 Escherichia coli BL2l (DE3)/pHCE-AST는 360 nm상에서 4-methylumbellifrrone에 의한 강한 형광을 보였고, 이는 대장균에서 arylsulfatase가 활성형으로 생산되었음을 의미하였다. E. coli BL21 (DE3)/pHCE-AST를 0.4% glycerol 또는 0.4% glucose가 포함된 LB 배지로 배양했을 때 arylsulfatase활성은 glycerol이 포함된 배지에서 활성이 더 높게 나타났다. 2% glycerol이 포함된 LB배지에서 arylsulfatase 활성은 약 15.0 unit/ml에 달했으며, 이는 1% glycerol을 첨가해서 배양했을 때보다 2.6배 이상의 높은 발현 수준이였다. 재조합 arylsulfatase 효소로 제조된 agarose와 시판용 agarose를 DNA markers를 이용해서 전기영동 성능을 비교했을 때 우수한 이동성과 분리능을 보였다. 본 연구의 결과, E. coli에서 과발현 생산된 arylsulfatase 효소를 이용하여 전기영동용 고순도 agarose생산 공정에 적용 가능함을 확인하였다.

한국인의 비소세포폐암종에서 O6-methylguanine-DNA methyltransferase (MGMT)의 발현도 분석 (Immunohistochemical Expression of O6-methylguanine-DNA Methyltransferase (MGMT) in Korean Patients with Non-Small Cell Lung Cancer.)

  • 이경은;홍영습;최필조;노미숙
    • 생명과학회지
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    • 제18권4호
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    • pp.580-584
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    • 2008
  • 본 연구에서는 손상된 DNA를 수복하는 중요한 효소로 알려진 $O^6-methylguanine-DNA$ methyltransferase (MGMT)발현의 의미를 비소세포폐암종에서 면역조직화학 염색법으로 알아보고자 하였다. 동아대학교 의료원에서 2001년부터 2004년까지 외과적으로 적출한 폐암종 조직 중 비소세포암종으로 진단된 74예를 연구대상으로 하였다. 면역염색 결과, MGMT 발현은 총 74예 중 49예(66.2%)에서 양성을 보였으며, 25예(33.8%)에서 단백 소실을 보였다. 조직학적 유형에 따른 결과를 살펴보면, 편평세포암종은 8/39예(20.5%)에서 단백 소실이 보였고, 샘암종은 17/35예(48.6%)에서 단백 소실이 관찰되어 통계적으로 유의한 차이가 관찰되었다(p=0.021). 하지만 나이, 성별, 흡연유무, 종양 크기, T 병기 및 림프절 전이에 따른 유의한 차이는 관찰되지 않았다(p>0.05). MGMT 단백 발현 소실은 특히 promoter 메틸화와 연관되어 종양에서 관찰된다고 알려져 있으므로, 향후 연구에서는 비소세포폐암종의 MGMT 단백 소실에 대한 임상적 의의를 밝히기 위하여 promoter 메틸화 연구가 추가적으로 수행되어져야 될 것으로 사료된다.

The Basis of Different Sensitivities of Ovarian Cancer Cells to the Recombinant Adenoviral Vector System Containing a Tumor-Specific L-plastin Promoter and E. coli Cytosine Deaminase Gene as a Transcription Unit

  • Chung, In-Jae
    • Biomolecules & Therapeutics
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    • 제17권2호
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    • pp.138-143
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    • 2009
  • We have reported previously on a replication incompetent recombinant adenoviral vector, AdLPCD, in which the expression of cytosine deaminase gene (CD) is driven by the tumor-specific L-plastin promoter. AdLPCD vector had been evaluated for its efficacy of chemosensitization of ovarian cancer cells to 5-FC. In spite of the fact that ovarian cancer cells, i.e., OVCAR-3 and SK-OV-3, are capable for adenoviral transduction judged by LacZ reporter gene analysis, two cell lines demonstrated quite different sensitivities toward AdLPCD/5-FC system. In OVCAR-3 cells, infection of AdLPCD followed by exposure to 5-FC resulted in the suppression of cell growth with statistical significance. On the other hand, SK-OV-3 cells were more resistant to the CD/5-FC strategy compared with OVCAR-3 cells under the same condition. The object of study was to investigate factors that would determine the sensitivity to AdLPCD/5-FC. We evaluated conversion rate of 5-FC to 5-FU after infection of AdLPCD by HPLC analysis, $IC_{50}$ of 5-FU, the expression level of integrin receptors i.e., ${\alpha}v{\beta}3$ and ${\alpha}v{\beta}5$, and status of p53 in OVCAR-3 and SK-OV-3 cells. The results indicated that OVCAR-3 cells have few favorable features compared with SK-OV-3 cells to be more effective to the AdLPCD/5-FC strategy; higher level of ${\alpha}v{\beta}5$ integrin, higher rate of conversion of 5-FC into 5-FC, and lower $IC_{50}$ of 5-FU. The results suggest that the replacement of 5-FU with CD/5-FC in combination chemotherapy would be less toxic and much greater cytotoxicity than the conventional combination chemotherapy in some patients.

법랑아세포종에서 p16과 E-cadherin의 메틸화 (Methylation of p16 and E-cadherin in ameloblastoma)

  • 박찬웅;윤혜경;박상준
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제36권6호
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    • pp.453-459
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    • 2010
  • Introduction: Ameloblastic carcinoma is a rare malignant lesion, and may arise from either carcinoma ex-ameloblastoma or de novo carcinoma. Aberrant promoter hypermethylation of the tumor-associated genes leading to their inactivation is a common event in many cancer types. The p16/CDKN2/INK4A gene and p16 5 protein are involved directly in regulating the cell cycles. Cadherins are cell adhesion molecules that modulate the epithelial phenotype and regulate tumor invasion. The aim of this study was to evaluate the roles of p16 and E-cadherin methylation and loss of p16 and E-cadherin expression in the malignant transformation of an ameloblastoma. Materials and Methods: Eight cases of ameloblastoma, including 4 benign ameloblastomas without recurrence, 2 benign ameloblastomas with recurrence and 2 carcinoma ex-ameloblastomas, were examined. The promoter hypermethylation profile of the p16 and E-cadherin genes was studied using methylation-specific polymerase chain reaction (MSP) and immunohistochemical staining for p16 and E-cadherin expression. Results: 1) Aberrant CpG island methylation of the p16 gene was detected in 3 of the 4 benign ameloblastomas without recurrence and 1 of the 2 benign ameloblastomas with recurrence. 2) Aberrant CpG island methylation of the E-cadherin gene was found in 1 of the 4 benign ameloblastomas without recurrence. 3) A loss of p16 expression was noted in 1 of 4 benign ameloblastomas without recurrence and 1 of 2 carcinoma ex-ameloblastomas. 4) A loss of E-cadherin expression was noted in 2 of the 4 benign ameloblastomas without recurrence, 1 of the 2 benign ameloblastomas with recurrence and 2 of the 2 carcinoma ex-ameloblastomas. 5) A loss of p16 expression was observed in 1 of the 4 cases showing aberrant methylation of the p16 gene. 6) A loss of E-cadherin expression was observed in 3 benign ameloblastoma case showing aberrant methylation of the E-cadherin gene. Conclusion: These results suggest that loss of E-cadherin expression related to the other genetic pathway (not methylation) might be an adjuvant indicator predicting the malignant transformation of an ameloblastoma. However, the number of samples in this study was too small and the relationship between the treatment methods and clinical course were not defined. Therefore, further study will be needed.

Fermentation and Purification of LacZ-Fused Single Chain Insulin Precursor for($B^{30}$-Homoserine) Human Insulin

  • SeungYup Lee;Jeo
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제1권1호
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    • pp.9-12
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    • 1996
  • In order to produce the single chain precursor of a novel human insulin analogue, (B30-Homoserine) insulin, the fermentative behaviors of Escherichia coli JM103 were studied, which harbors pKBA plasmid carrying a hybrid gene in which the gene for a single chain precursor was fused with lacZ gene under tac promoter. The maximal induction of gene expression was achieved when more than 0.05 mM of isopropyl-$\beta$-D-thiogalactopyranoside(IPTG) was supplemented to fermentation medium after 4 h cultivation of E. coli, and followed by longer than 2-h fermentation. The hybrid protein of the single chain insulin precursor was isolated from cytoplasmic inclusion bodies by dissolving in 8M urea solution, and purified through DEAE-Sephacel and Sephadex G-200 column chromatographies with a recovery of 35%. The finally purified hybrid protein showed a single band on sodium dodecyl sulfate-polyacrylamide gel.

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