• Title/Summary/Keyword: M/BP/1

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Agrobacterium-Mediated Genetic Transformation of Pepper for the Development of Blight Resistant Cultivar (고추의 역병 저항성 품종 개발을 위하여 Agrobacterium tumefaciens를 이용한 elicitin 유전자 도입)

  • Kwon, Tae-Ryong;Lee, Moon-Jung;Han, Jung-Sul;Shin, Dong-Hyun;Oh, Jung-Youl;Kim, Kyung-Min;Kim, Chang-Kil
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.55-59
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    • 2007
  • The study was carried out to develop transformants resisting to Phyophthora blight disease in the domestic pepper cultivar Subicho. In transforming of syn600 promoter with elicitin gene using Agrobacterium (LBA4404/pBI101 syn600-syn${\alpha}$-elicitin) to cotyledons of pepper, rate of shoot formation in 'Subicho' was 11.1% in medium containing 3 mg/L zeatin and 0.05 mg/L NAA, and also 12.8% in medium containing combination of 4 mg/L zeatin and 0.05 mg/L MAA. For PCR reaction using elicitin gene primer of transformants regenerated from cotyledons, we detected a specific band of 536 bp, and also showed strong signal at position of 536 bp in accordance with NPTII gene used as probe in Southern blot. Transformants pepper shown resistance to blight fungus was inoculated to seedlings of the $T_{1}\;and\;T_{2}$ transformants by concentration (density: zoo spore $10^{3}/mL$).

Ceramide Induces Apoptosis and Growth Arrest of Human Glioblastoma Cells by Inhibiting Akt Signaling Pathways

  • Lee, Eun-Chang;Lee, Young-Seok;Park, Na-Hee;So, Kwang-Sup;Chun, Young-Jin;Kim, Mie-Young
    • Biomolecules & Therapeutics
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    • v.19 no.1
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    • pp.21-26
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    • 2011
  • Ceramide is an important lipid mediator of extracellular signals that control various cellular functions, including apoptosis. In this study, we showed that ceramide induced apoptosis in U373MG human glioblastoma cells associated with G1 cell cycle arrest. Treatment of cells with ceramide increased proapoptotic Bax expression and inhibited the expression of antiapoptotic Bcl-2 and Bcl-xL Ceramide also downregulated cyclin E, cyclin D1, cdk 2, and cdk4 which are involved in regulating cell cycle. In addition, ceramide suppressed phosphorylation of Akt, Bad, p70 S6 kinase, and 4E-BP1, suggesting the involvement of Akt/mTOR signaling pathway. Additionally, okadaic acid, an inhibitor of protein phosphatase 2A, partially blocked the ceramide mediated inhibition of phosphorylation of Akt and 4E-BP1. These results suggest that ceramide induces apoptosis in U373MG glioblastoma cells by regulating multiple signaling pathways that involve cell cycle arrest associated with Akt signaling pathway.

Cloning and Expression Characteristics of Pharbitis nil COP1 (PnCOP1) During the Floral Induction

  • Kim, Yun-Hee;Kim, Seong-Ryong;Heo, Yoon-Kang
    • Journal of Photoscience
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    • v.12 no.1
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    • pp.1-9
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    • 2005
  • The ubiquitin E3 ligase COP1 (Constitutive Photomorphogenesis 1) is a protein repressor of photomorphogenesis in Arabidopsisplants, and it found in various organisms, including animals. The COP1 protein regulates the stability of many of the light-signaling components that are involved in photomorphogenesis and in the developmental processes. To study the effect of COP1 on flowering in a short day plant, we have cloned a full-length of PnCOP1 (Pharbitis nil COP1) cDNA from Pharbitis nil Choisy cv. Violet, and we examined its transcript levels under various conditions. A full-length PnCOP1 cDNA consists of 2,280 bp nucleotidesthat contain 47 bp of 5'-UTR, 232 bp of 3'-UTR including the poly (A) tail, and 1,998 bp of the coding sequence. The deduced amino acid sequence contains 666 amino acids, giving it a theoretical molecular weight of 75 kD and a isolectric point of 6.2. The PnCOP1 contains three distinct domains, an N-terminal $Zn^2+$-binding RING-finger domain, a coiled-coil structure, and WD40 repeats at the C-terminal, implying that the protein plays a role in protein-protein interactions. The PnCOP1 transcript was detected in the cotyledon, hypocotyls and leaves, but not in root. The levels of the PnCOP1 transcript were reduced in leaves that were a farther distance away from the cotyledons. The expression level of the PnCOP1 gene was inhibited by light, while the expression was increased in the dark. During the floral inductive 16 hour-dark period for Pharbitis nil, the expression was increased and it reached its maximum at the 12th hour of the dark period. The levels of PnCOP1 mRNA were dramatically reduced upon light illumination. These results suggest that PnCOP1 may play an important function in the floral induction of Pharbitis nil.

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Characterization of Ecdysteroid UDP-Glucosyltransferase Gene Promoter from Bombyx mori Nucleopolyhedrovirus

  • Zhang, Zhi-Fang;Shen, Xing-Jia;Yi, Yong-Zhu;Tang, Shun-Ming;Li, Yi-Ren;He, Jia-Lu;Wu, Xiang-Fu
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.2
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    • pp.169-174
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    • 2004
  • Bombyx mori nucleopolyhedrovirus(BmNPV) ecdysteroid UDP-glucosyltransferase gene (egt) promoter fragments of different lengths were amplified from BmNPV ZJ-8 genomic DNA by PCR. Reporter plasmids pBmegt542-luc, pBmegt309-luc and pBmegtl59-luc with luciferase (lue) driven by egt promoters were constructed. Both in vitro and in vivo expressions showed that BmNPV egt promoter activity requires the transactivation of viral factor(s), and expression of luc was detected earliest at 24 hrs post infection (pi). BmNPV ZJ-8 homologous region 3 (hr3) increased the expression of luc by over 1,600-fold. Molting hormone of 1.0 - 2.0 $\mu\textrm{g}$/$m\ell$ can dramatically down regulate expression of luc. Juvenile hormone analogue of 0.5-2.0 ${\mu}g$/$m\ell$ increased expression of luc by 145.8% to 75.7%. Deletion assay revealed that the promoter fragment of 159 bp contains the basal promoter structure; Promoter fragments of 309 bp and 542 bp showed similar but much higher transcriptional activities than that of 159 bp, suggesting that nucleotide from -159 to -309 nt upstream the translation initiation site harbors the main cis-acting elements.

Study on 2BP Distance Bounding Protocol to Prevent Relay Attack in RFID environment (RFID 환경에서 중계공격을 방지를 위한 2BP Distance Bounding 프로토콜 연구)

  • Jung, Youn-Sung;Kim, Hyeong-Ju;Lee, A-Young;Jun, Moon-Seog
    • Proceedings of the Korea Information Processing Society Conference
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    • 2012.11a
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    • pp.892-895
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    • 2012
  • RIFD 시스템은 개체 인증, 전자결제 등 다양한 분야에서 사용되고 있다. 무선통신의 특성상 악의적인 공격자로부터 중계공격이 가능하여 공격자가 정당한 사용자로 가장할 수 있다. 이러한 중계공격을 예방하기 위하여 Distance Bounding이라는 개념의 프로토콜이 제안되어 왔다. 하지만, 기존 연구는 태그 ID 전달의 기밀성을 보장하지만 ID 검색의 비효율성 및 불필요한 두 번째 저속 단계가 존재하는 단점이 있다. 따라서 본 논문은 ID의 검색 없고, 시도 값과 응답 값을 2bits로 하여 마지막 저속 단계를 생략하며, 고속 단계에서 공격자의 공격 탐지 및 Terrorist 공격에 대하여 $(1/2)^n$의 공격 성공률을 갖는 2BP(2Bit 2Phase) Distance Bounding 프로토콜을 제안한다.

The Cytotoxic Constituents of Betula platyphylla and their Effects on Human Lung A549 Cancer Cells

  • Yang, Eun-Ju;An, Ju-Hee;Son, Youn Kyoung;Yeo, Joo-Hong;Song, Kyung-Sik
    • Natural Product Sciences
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    • v.24 no.4
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    • pp.219-224
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    • 2018
  • During the screening for cytotoxic compounds from plants grown in Korea, Betula platyphylla (BP) showed potent activity against the adenocarcinomic human alveolar basal epithelial A549 cell line. To identify the cytotoxic components from BP, the $CH_2Cl_2$ fraction with the most significant cytotoxic effect was applied to the column chromatographies. Seven compounds were isolated: lupeol (1), betulinic acid (2), (-)-rhododendrol (3), platyphyllenone (4), platyphyllone (5), (-)-centrolobol (6), and oleanolic acid (7). Among them, three diarylheptanoids (4 - 6) exhibited cytotoxicity toward A549 cells. Especially, $50{\mu}M$ of 4 reduced A549 cell viability to $18.93{\pm}0.82%$ compared to control ($100.00{\pm}21.48%$). Lactate dehydrogenase (LDH) leakage and intracellular reactive oxygen species (ROS) production were also induced by $50{\mu}M$ 4. This is the first report on the cytotoxic effect of BP-derived diarylheptanoids 4-6 against A549 cells. The compound 4 may be useful for the development of early hit compounds for non-small cell lung carcinoma, but the consideration about selectivity of 4 is required since 4 also showed the cytotoxicity in the human normal lung epithelial BEAS-2B cell line.

Expressions of Pituitary Adenylate Cyclase-Activating Polypeptide and Its Receptor Gene in the Rat Uterus (흰쥐 자궁에서 Pituitary Adenylate Cyclase-Activating Polypeptide와 수용체 유전자의 발현)

  • 이성호
    • Development and Reproduction
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    • v.2 no.1
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    • pp.21-27
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    • 1998
  • The present study was performed to analyze the gene expressions of pituitary adenylate cyclase-activating polypeptide(PACAP) and its receptor in the rat uterus, a candidate for novel extrahypothalamic source and target. The PACAP cDNA fragments corresponding to the common exon region which is found in both the rat hypothalamus and testis were produced from all tissue samples including the rat uterus by reverse transcriptionpolymerase chain reaction (RT-PCR). No PCR product was amplified from the rat hypothalamic, pituitary, ovarian and uterine samples when the 5' primer corresponding to the testis-specific exon 1 region was used, while the predicted size of product was detected from the testis sample. RT-PCR using the uterine RNA and specific primers for the PACAP receptor yielded products with predicted sizes. Transcripts for the rat uterine PACAP receptor were identified as type I isoforms with hip-hop and hip- or hop-type inserts. After pregnant mare's serum gonadotropin (15 IU) treatment of immature rats (day 25), the level of PACAP mRNA was increased in 24 h and 48 h group, and was declined to the lowest in 72 h group. The present study shows the presence of transcripts for PACAP and its receptor isoform in the rat uterus. These finding ssuggest that the uterine PACAP ight act as a novel autocrine and/or paracrine factor via its specific receptors on the reglulation of rat uterine function and physiology during the reproductive cycle.

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Anti-Proliferation Effects and Molecular Mechanisms of Action of Tetramethypyrazine on Human SGC-7901 Gastric Carcinoma Cells

  • Ji, Ai-Jun;Liu, Sheng-Lin;Ju, Wen-Zheng;Huang, Xin-En
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.8
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    • pp.3581-3586
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    • 2014
  • Aim: To investigate the effects of tetramethypyrazine (TMP) on proliferation and apoptosis of the human gastric carcinoma cell line 7901 and its possible mechanism of action. Methods: The viability of TMP-treated 7901 cells was measured with a 3-(4, 5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT) and cell apoptosis was analyzed by flow cytometry. The distribution of cells in different phases of cell cycle after exposure of TMPs was analyzed with flow cytometry. To investigate the molecular mechanisms of TMP-mediated apoptosis, the expression of NF-${\kappa}Bp65$, cyclinD1 and p16 in SGC-7901 cells was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and western blotting. Results: TMP inhibited the proliferation of human gastric carcinoma cell line 7901 in dose and time dependent manners. Cell growth was suppressed by TMP at different concentrations (0.25, 0.5, 1.0, 2.0 mg/ml), the inhibition rate is 0.46%, 4.36%, 14.8%, 76.1% (48h) and 15.5%, 18.5%, 41.2%, 89.8% (72h) respectively. When the concentration of TMPs was 2.0mg/ml, G1-phase arrest in the SGC-7901 cells was significant based on the data for cell cycle distribution. RT-PCR demonstrated that NF-${\kappa}Bp65$ and cyclin D1 mRNA expression was significantly down-regulated in 7901 cells treated with 2.0 mg/ml TMP for 72h (p<0.05), while the p16 mRNA level was up-regulated (p<0.05). The protein expression of NF-${\kappa}Bp65$ and cyclin D1 decreased gradually with the increase in TMP concentration, compared with control cells (p<0.05), while expression of protein p16 was up-regulated (p<0.01). Conclusion: TMP exhibits significant anti-proliferative and pro-apoptotic effects on the human gastric carcinoma cell line SGC-7901. NF-${\kappa}Bp65$, cyclinD1 and p16 may also play important roles in the regulation mechanisms.

Synthesis and Physical Properties of Liquid-Crystalline Polyurethanes (液晶性 Polyurethane의 合成과 物性에 關한 硏究)

  • Lee, Jong Back;Song, Jin Cherl;Choi, Dae Woong
    • Textile Coloration and Finishing
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    • v.8 no.1
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    • pp.56-63
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    • 1996
  • A number of thermotropic liquid crystalline polyurethanes with mesogenic unit were synthesized by polyaddition of a para-type diisocyanate such as 1, 4-phenylene diisocyanate(1,4-PDI) with 4, 4'-bis($\omega$-hydorxyalkoxy) biphenyls($BP{m}$) in DMF. The thermal and liquid crystab line properties were examined by differential scanning calorimetry(DSC), polarized optical microscopy, and wide-angle X-ray scattering(WAXS). Intrinsic viscosities of the polymers exbibited two endothermic peaks correspondinding to phase transitions of melting and isotropization. For example, polyurethane(1,4-PDI/($BP{11}$) ) was found to display a liquid crystalline phase between 177 to 205$^{\circ}C$. In order to know how the hydrogen bonding interaction affects the formation of mesophases in polyurethane 1, 4-PDI/($BP{8}$) / thermal processing FT-IR measurements were carried out. It was found that the stretches regarded as shift to higher frequency region with increasing temperature which showed grdually their liquid crystalline phase

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mTOR Signal Transduction Pathways Contribute to TN-C FNIII A1 Overexpression by Mechanical Stress in Osteosarcoma Cells

  • Zheng, Lianhe;Zhang, Dianzhong;Zhang, Yunfei;Wen, Yanhua;Wang, Yucai
    • Molecules and Cells
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    • v.37 no.2
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    • pp.118-125
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    • 2014
  • Osteosarcoma is the most common primary malignant bone tumor with a very poor prognosis. Treating osteosarcoma remains a challenge due to its high transitivity. Tenascin-C, with large molecular weight variants including different combinations of its alternative spliced FNIII repeats, is specifically over expressed in tumor tissues. This study examined the expression of Tenascin-C FNIIIA1 in osteosarcoma tissues, and estimated the effect of mechanical stimulation on A1 expression in MG-63 cells. Through immunohistochemical analysis, we found that the A1 protein was expressed at a higher level in osteosarcoma tissues than in adjacent normal tissues. By cell migration assay, we observed that there was a significant correlation between A1 expression and MG-63 cell migration. The relation is that Tenascin-C FNIIIA1 can promote MG-63 cell migration. According to our further study into the effect of mechanical stimulation on A1 expression in MG-63 cells, the mRNA and protein levels of A1 were significantly up-regulated under mechanical stress with the mTOR molecule proving indispensable. Meanwhile, 4E-BP1 and S6K1 (downstream molecule of mTOR) are necessary for A1 normal expression in MG-63 cells whether or not mechanical stress has been encountered. We found that Tenascin-C FNIIIA1 is over-expressed in osteosar-coma tissues and can promote MG-63 cell migration. Furthermore, mechanical stress can facilitate MG-63 cell migration though facilitating A1 overexpression with the necessary molecules (mTOR, 4E-BP1 and S6K1). In con-clusion, high expression of A1 may promote the meta-stasis of osteosarcoma by facilitating MG-63 cell migration. Tenascin-C FNIIIA1 could be used as an indicator in metastatic osteosarcoma patients.