• Title/Summary/Keyword: agarose gel

Search Result 429, Processing Time 0.033 seconds

Apoptosis in Human Corneal Epithelial cells induced by Exhausted Medium (Exhausted Medium에 의한 각막상피 세포의 세포고사 유도)

  • Kim, Jae-Min
    • Journal of Korean Ophthalmic Optics Society
    • /
    • v.5 no.1
    • /
    • pp.83-87
    • /
    • 2000
  • To investigate exhausted-medium-induced apoptosis in human corneal epithelial(HCE) cells, this study was performed DNA gel electrophoresis, M30 CytoDEATH staining and FAS-FAS ligand ELISA. SV-40 transfected cells were grown to confluency in culture for 7days. The supernatant was harvested and filtered with $0.22{\mu}m$ filter paper. Fresh HCE cells were exposed to the filtered exhausted medium for 1~2 days. Apoptotic cells were prepared for DNA extraction and run the agarose gel for DNA ladder pattern. M30 CytoDEATH was used a tool for easy and reliable determination of very early apoptosis in HCE cells. The control and exhausted medium were assayed for soluble FAS/FAS ligand protein by ELISA. HCE cells exposed to exhausted medium showed a typical DNA ladder pattern. Sporadic M30 CytoDEATH positive cells were detected among HCE cells exposed to exhausted medium. Soluble FAS/FAS ligand levels were not elevated in the exhausted medium compared to the fresh medium control. This study suggests that possible mechanism of exhausted medium induced apoptosis does not include the FAS-FAS ligand system.

  • PDF

In-Vivo Heat Transfer Measurement using Proton Resonance Frequency Method of Magnetic Resonance Imaging (자기 공명영상 시스템의 수소원자 공명 주파수법을 이용한 생체 내 열 전달 관찰)

  • 조지연;조종운;이현용;신운재;은충기;문치웅
    • Journal of the Institute of Electronics Engineers of Korea SC
    • /
    • v.40 no.3
    • /
    • pp.172-180
    • /
    • 2003
  • The purpose of this study is to observe the heat transfer process in in-vivo human muscle based on Proton Resonance Frequency(PRF) method in Magnetic Resonance Imaging(MRI). MRI was obtained to measure the temperature variation according to the heat transfer in phantom and in-vivo human calf muscle. A phantom(2% agarose gel) was used in this experiment. MR temperature measurement was compared with the direct temperature measurement using a T-type thermocouple. After heating agarose gel to more than 5$0^{\circ}C$ in boiling hot water, raw data were acquired every 3 minutes during one hour cooling period for a phantom case. For human study heat was forced to deliver into volunteer's calf muscle using hot pack. Reference data were once acquired before a hot pack emits heat and raw data were acquired every 2 minutes during 30minutes. Acquired raw data were reconstructed to phase-difference images with reference image to observe the temperature change. Phase-difference of the phantom was linearly proportional to the temperature change in the range of 34.2$^{\circ}C$ and 50.2$^{\circ}C$. Temperature resolution was 0.0457 radian /$^{\circ}C$(0.0038 ppm/$^{\circ}C$) in phantom case. In vivo-case, mean phase-difference in near region from the hot pack is smaller than that in far region. Different temperature distribution was observed in proportion to a distance from heat source.

Quantitative Counting of Bifidobacterium spp. in a Sample Mixed with Lactobacillus acidophilus

  • Park, Young-Min;So, Jae-Seong
    • Journal of Microbiology and Biotechnology
    • /
    • v.8 no.2
    • /
    • pp.182-184
    • /
    • 1998
  • PCR was used for quantitative counting of Bifidobacterium spp. in a sample mixed with Lactobacillus acidophilus using two primer sets; one set for universal priming and the other set for Bifidobacterium specific priming. DNA products from two independent PCRs with DNA extracted from the mixed sample were found to be easily distinguishable from each other by agarose gel electrophoresis. The concentrations of PCR products correlated with the total number of bacteria and with the number of Bifidobacterium spp. present in the sample.

  • PDF

Antioxidative Effects of Cichorium intybus Root Extract on LDL (Low Density Lipoprotein) Oxidation

  • Kim, Tae-Woong;Yang, Ki-Sook
    • Archives of Pharmacal Research
    • /
    • v.24 no.5
    • /
    • pp.431-436
    • /
    • 2001
  • The water extract of Cichorium intybus (WECI) showed a remarkable antioxidative effect on LDL, and inhibitory effects on the production of thiobarbituric acid reactive substance and the Degradation of fatty acids in LDL. Vitamin 1 and unsaturated fatty acids in LDL were protected by adding WECI from the effects of metal catalyzed LDL oxidation. From the results obtained, we conclude that LDL oxidation is inhibited in vitro by the addition of WECI, and that LDL is protected by WECI from oxidative attack, as shown by agarose gel electrohporesis.

  • PDF

Codonoposide $1_c$ is a potent inducer of apoptosisin Human Leukemia cell line, HL-60.

  • Lee, Kyung-Won;Lee, Kyung-Tae;Park, Hee-Juhn;Choi, Jong-Won
    • Proceedings of the PSK Conference
    • /
    • 2003.10b
    • /
    • pp.159.2-159.2
    • /
    • 2003
  • Codonopside lc is an active natural compound isolated from the roots of Codonopsis lanceolata (Campanulaceae), a Korean medicinal herb. In the present study, we investigated the in vitro effect of Codonopside 1c on the proliferation and induction of apoptosis in HL-60 human promyelocytic cells. When HL-60 cells were treated with Codonopside 1c, evidence of apoptotic features, including DNA fragmentation, formation of DNA ladder in agarose gel elecrophoresis and increse of annexin V binding, were obtained. (omitted)

  • PDF

Purification and Cellular Localization of Extracellular Nuclease of Serratia marcescens Expressed in Escherichia coli (대장균에 발현된 Serratia marcescens의 Nuclease의 정제와 세포내 분포)

  • Kim, Woe-Yeon;Lee, Hoon-Sil;Suh, Sook-Jae;Cho, Moo-Je;Lee, Sang-Yeol;Kim, Jae-Won
    • Korean Journal of Microbiology
    • /
    • v.32 no.2
    • /
    • pp.147-154
    • /
    • 1994
  • Nuclease was secreted to the environmental media from the Escherichia coli JM107 tranformant harboring the extracellular nuclease gene of Serratia marcescens in the plasmid of pNUC4. Under the growth conditions, the amount of secreted enzyme was increased in parallel with bacterial growth conditions, the amount of secreted enzyme was increased in parallel with bacterial growth. The enzyme was purified using chromatofraphic procedures of Matrex green gel and heparin agarose affinity gel, resulted in 50-fold purification with 15% recovery of the enzyme. The apparent molecular weight of the enzyme was estimated to be 29Kda by sodium dodecylsulfate denaturing gel electrophoresis. Using the purified enzyme, polyclonal antibody was obtained from the rabbit. The specificity of the antibody was confirmed by immunoblotting and immunoprecipitaion. For the investigation of cellular distribution of the enzyme, cells were fractionated into three fractions; cytoplasm, periplasm and extracellular fluid. While more than 80% of the enzymatic activity was detected in the extracellular fluid and periplasm, a little was found in the cytoplasm, indicating that the enzyme was likely to be immediately exported to the membrane for excretion after biosynthesis. These results were confirmed again by immunocytochemistry technique using the antibody.

  • PDF

Determinant Involved in the Loss of Pathogenicity in Wilt - Inducing Pseudomonas solanacerum (마름병 병원균 Pseudomonas solanacearum의 병원성 상실요인에 관하여)

  • 김을제;윤경란;이영하;이청호;박지창;최광태
    • Journal of the Korean Society of Tobacco Science
    • /
    • v.12 no.1
    • /
    • pp.9-18
    • /
    • 1990
  • To study the determinants which are involved in the loss of pathogenicity in wilt-inducing Pseudomoms solamcewum, several physlologica I functions were compared in a virulent P. solanacearum strain and an avirulent, spontaneously derived mutant strain. The polyacrylamide gel electrophoresis showed the distinction between two strains in the patterns and the relative intensity of proteins produced intracellularly or extracellularly. Enzyme assays showed that the level of polygalacturonase activity in the culture filtrate of the avirulent mutant was markedly reduced, while carboxymethylcellulase(rondoglucanase) activity in both strains were nearly negligible. These results suggest that the loss of pathogenicity in mutant strain is attributed in part to the reduced production of polygalacturonase. In audit ion, comparative analyses by agarose gel electrophoresis of DNA molecules isolated from both strains show that the pathogenicity genes of p. solanaceerum are not located on plasmid but are on chromosome.

  • PDF

Megabase-sized DNA isolation and electrophoretic karyotype of fusarium oxysporum schlecht

  • Park, Min-Seon;Min, Byung-Re
    • Journal of Microbiology
    • /
    • v.33 no.2
    • /
    • pp.132-135
    • /
    • 1995
  • To investigate the electrophoretic karytype of Fusarium oxysporum, intact chromosomal DNA was separated by pulsed-field gel electrophoresis (PEGE). DNA extraction from nulcei, mycelia and protoplasts were compared with one another and with the quantity and the suitability for PFGE separation in agarose gel. As a result, the most useful extracting method for intact DNA was found to be that from protoplasts. By varying the electrophoretic conditions, 8 chromosomal DNA bounds were resolved. Using the Schizosaccharomyces pombe and Saccharomyces cerevisiae as size standards, the size of Fusarium oxysporum chromosomes was estimated to range from approximately 0.6 Mb TO 6.7 Mb, and total genome size was 26.7 Mb. The suitability of electrophoretic karyotyping as a tool for strain characterization is discussed.

  • PDF

A Second Thioltransferase of Schizosaccharomyces pombe Contains Glutathione S-transferase Activity

  • Kim, Hong-Gyum;Park, Eun-Hee;Lim, Chang-Jin
    • BMB Reports
    • /
    • v.32 no.6
    • /
    • pp.535-540
    • /
    • 1999
  • Two types of the thioltransferase (also called glutaredoxin) have been previously detected in the cytosolic extract of Schizosaccharomyces pombe, a fission yeast. Previously, the one with a smaller molecular mass (14kDa) was purified and characterized. In the present study, the second thioltransferase was purified. The purification procedure included ammonium sulfate fractionation (40-80%), Sephadex G-200 gel filtration, DEAE-cellulose ion-exchange chromatography, Sephadex G-50 gel filtration, and glutathione-agarose affinity chromatography. The purified enzyme showed a single band on SDS-PAGE, and its molecular mass was determined to be 23 kDa. It utilizes various compounds as substrates, including 2-hydroxyethyl disulfide. Interestingly, we found that the purified thioltransferase also contains significant glutathione S-transferase activity.

  • PDF

Cloning of a Alkaline Protease Gene from Xanthomonas sp. YL-37 (Xanthomonas sp. YL-37의 Alkaline Protease 유전자의 클로닝)

  • 이대희;김수경;이승철;윤병대;황용일
    • Microbiology and Biotechnology Letters
    • /
    • v.23 no.2
    • /
    • pp.145-149
    • /
    • 1995
  • For the purpose of developing a new biodegradable detergent, we have isolated a gene encoding wide-range temperature applicable alkaline protease from Xanthomonas sp. YL-37 (Lee et al., 1994, Kor. J. Appl. Microbiol. Biotechnol.). An alkaline protease gene was isolated from the gene bank that was prepared from the chromosomal DNA of Xanthomonas sp. YL-37. From the results of agarose gel electrophoresis and a restriction enzyme mapping, a 2.7 kb DNA fragment containing the alkaline protease gene was inserted in the plasmid pUC9. Extracellular activity of a clone having alkaline protease gene was detected on SDS-polyacrylamide gel with activity staining assay. The molecular weight of alkaline protease was determined to be about 64 kDa from 11% SDS-PAGE analysis. Alkaline protease activity, produced from E. coli which harboring the plasmid, showed no difference at reaction temperature 20, 30 and 40$\circ$C, respectively. This result showed that alkaline protease produced from E. coli harboring the plasmid was apparently the same as that of Xanthomonas sp. YL-37.

  • PDF