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분자유전학적인 기술을 이용한 육 감별법

  • Kim, Tae-Heon
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2000.11a
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    • pp.59-75
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    • 2000
  • This study was carried out to develop a DNA marker for identifying between Korean cattle (Hanwoo) and other breeds. First experiment was performed to isolate Hanwoo specific DNA marker at sequence characterized amplified regions (SCARs). Five breeds of cattle including Hanwoo, Holstein, Hereford, Angus and Charolais were represented with the from 8 to 20 individuals. Fourteen primers of 300 arbitrary primers of 10 nucleotides showed reproducible polymorphism across the breeds. An amplified band of 0.9 kb in the primer MG-3 showed the specificity to Holstein breed. And MG-6 and MG-12 detected the Hereford and Hanwoo specific markers at the size of 2.0 kb and 1.0 kb, respectively. A 1.0 kb band of MG-12 was cloned and sequenced. A SCAR primer was designed based on the obtained sequences. It was possible to identify the Hanwoo from Holstein breed. Second experiment was carried out to observe the genotype frequencies of MC1R in 1,044 samples of imported beef and eight different cattle breeds including Hanwoo, Holstein, Angus, Brown-Swiss, Charolais, Limousin, Simmental and Hereford. The primers for the amplification of bovine MC1R gene were designed based on a bovine MC1R gene sequence (GenBank accession no.Y19103). A size of 350 bp was amplified by polymerase chain reaction(PCR), digested with two different restriction enzyme, BsrFI and MspA II, and electrophoresed in 2.5% Metaphore agarose gel for determination of genotypes. Genotype frequencies of Hanwoo were 0.10 in E+e and 0.90 in ee. Allele ED was shown in all of Holstein and Angus breeds tested which have black coat color phenotypes. We suggested that SCAR marker and the bovine MC1R gene could be used as a DNA marker for distinguishing beef between Hanwoo and Holstein.

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Preparation of Polymer Gel Electrolyte for EDLCs using P(VdF-co-HFP)/PVP (P(VdF-co-HFP)/PVP를 이용한 EDLC용 고분자 겔 전해질의 제조)

  • Jung, Hyun-Chul;Jang, In-Young;Kang, An-Soo
    • Applied Chemistry for Engineering
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    • v.17 no.3
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    • pp.243-249
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    • 2006
  • Porous polymer gel electrolytes (PGEs) based on poly(vinylidenefluoride-co-hexafluoropropylene) (P(VdF-co-HFP)) as a polymer matrix and polyvinylpyrolidone (PVP) as a pore-forming agent were prepared and electrochemical properties were investigated for an electric double layer capacitor (EDLC) in order to increase a permeability of an electrolyte into the PGE. Propylene carbonate (PC) and ethylene carbonate (EC) as plasticizers, and tetraethylammonium tetrafluoroborate ($TEABF_4$) as a supporting salt for the PGE were used. EDLC unit cells were assembled with the PGE and electrode comprising BP-20 and MSP-20 as activated carbon powders, Super P as a conducting agent, and P(VdF-co-HFP)/PVP as a mixed binder. Ion conductivity of PGEs increased with an increased PVP content and was the best at 7 wt% PVP, whereas electrochemical characteristics such as AC-ESR of unit cell were better in 3 wt%. And electrochemical characteristics of the unit cell with PGE were the best at a 33 : 33 weight ratio of PC to EC. Specific capacitance of a mixed plasticizer system of PE and EC was higher than that of pure PC. Ion conductivity of PGEs with a film thickness of $20{\mu}m$ was higher, but electrochemical characteristics of unit cells were higher for a $50{\mu}m$ membrane thickness. Also, the unit cell has shown the highest capacitance of 31.41 F/g and more stable electrochemical performance when PGE and electrode were hot pressed. Consequently, the optimum composition ratio of PGE for EDLCs was 23 : 66 : 11 wt% such as P(VdF-co-HFP) : PVP = 20 : 3 wt% and PC : EC = 44 : 22 wt%. In this case, $3.17{\times}10^{-3}S/cm$ of ion conductivity was achieved at the $50{\mu}m$ thickness of PGE for EDLCs. And the electrochemical characteristics of unit cells were $2.69{\Omega}$ of DC-ESR, 28 F/g of specific capacitance, and 100% of coulombic efficiency.

Methylation of O6-Methyl Guanine Methyltransferase Gene Promoter in Meningiomas - Comparison between Tumor Grades I, II, and III

  • Larijani, Leila;Madjd, Zahra;Samadikuchaksaraei, Ali;Younespour, Shima;Zham, Hanieh;Rakhshan, Azadeh;Mohammadi, Foruzan;Rahbari, Ali;Moradi, Afshin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.33-38
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    • 2014
  • Background: Meningiomas are the second most common primary intracranial tumors after gliomas. Epigenetic biomarkers such as DNA methylation, which is found in many tumors and is thus important in tumorigenesis can help diagnose meningiomas and predict response to adjuvant chemotherapy. We investigated aberrant O6-methyl guanine methyltransferase (MGMT) methylation in meningiomas. Materials and Methods: Sixty-one patients were classified according to the WHO grading, and MGMT promoter methylation status was examined via the methylation-Specific PCR(MSP) method. Results: MGMT promoter methylation was found in 22.2% of grade I, 35% of grade I with atypical features, 36% of grade II, and 42.9% of grade III tumors. Conclusions: There was an increase, albeit not statistically significant, in MGMT methylation with a rise in the tumor grade. Higher methylation levels were also observed in the male gender.

THE HYPERMETHYLATION OF E-CADHERIN GENE IN ORAL SQUAMOUS CELL CARCINOMA (구강 편평세포암에서 E-cadherin 유전자의 과메틸화)

  • Pyo, Sung-Woon;Kim, Young-Sill;Park, Ji-Young;Kim, Chang-Hyen;Lee, Won;Park, Min-Kyu
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.2
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    • pp.135-140
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    • 2008
  • Loss of E-cadherin (E-cad) expression has been found in multiple cancers and is postulated to facilitate tumor cell dissociation and metastais. Promotor methylation may provides an alternative pathway for loss of gene function. This study evaluated the role of hypermethylation in the down-regulation of E-cad in oral squamous cell carcinoma (OSCC). We examined the E-cad expression by immunohistochemical staining and detected methylation status by methylation-specific polymerase chain reaction (MSP) in 20 OSCC tissues. Overally, 12 (60%) cases of hypermethylation of E-cad were detected and we found there were no correlation between methylation and age, histologic grade, lympn node metastasis, tumor size and clinical stage. However, Eleven (73.3%) of 15 samples which was negative for E-cad staining showed hypermethylation of E-cad promotor region. On the other hand, only one (20%) of 5 E-cad positive sample was observed with methylated status. The underexpression of E-cad was found to be related to promotor hypermethylation (p=0.035). In conclusion, we suggest that hypermethylation play a role in inactivation of E-cad gene and may be a appreciable biomarker for diagnosis and treatment of OSCC.

Association Study of Dopamine Transporter(DAT1) G2319A Genetic Polymorphism in Alcohol Dependence (알코올의존 환자의 도파민 수송체(DAT1)G2319A의 유전자 다형성 연합연구)

  • Yang, Byung-Hwan;Lee, Mi-Gyung;Choi, Ju-Yoen;Oh, Dong-Yul;Kim, Gil-Sook;Kim, Hyung-Tae;Chai, Young-Gyu
    • Korean Journal of Biological Psychiatry
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    • v.8 no.2
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    • pp.239-245
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    • 2001
  • Objective : Dopamine transporter is member of family of Na/Cl dependent neurotransmitter transporter, 12 transmembrane domain, that has high substrate specificity, affinity. It is related with dopamine reuptake in presynaptic vesicle. DAT has a VNTR in its 3'-untranslated region(UTR). 3'-UTR VNTR polymorphism is related with modification of dopamine transmission. The association between with VNTR polymorphism and neuropsychiatric disorders such as alcohol dependence, and low activity ALDH has been studied, but their relationship is unclear. We study about association of 3'-UTR VNTR of DAT gene and G2319A and alcohol dependence. Method : Group of Korean subjects were studied with alcohol dependence(n=49 male) compared to mentally healthy controls(n=53 male). The peripheral blood sample was acquired, and Polymerase Chain Reaction(PCR) amplification, MspI procedure was done. Result : There was a significant difference between alcohol dependence group and normal control(genotype frequency p<0.05, allele frequency p<0.05) Allele A frequency and genotype(GG, GA) frequency was a significant difference between alcohol dependence group and normal control(p<0.05). Conclusion : Our study showed that genetic polymorphism of DAT1 G2319A had relation with alcohol dependence.

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Halotolerant Spore-Forming Gram-Positive Bacterial Diversity Associated with Blutaparon portulacoides (St. Hill.) Mears, a Pioneer Species in Brazilian Coastal Dunes

  • Barbosa Deyvison Clacino;Irene Von Der Weid;Vaisman Natalie;Seldin Lucy
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.193-199
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    • 2006
  • Halotolerant spore-forming Gram-positive bacteria were isolated from the root, rhizosphere, and non-rhizosphere soil of Blutaparon portulacoides. The different isolates were characterized genetically using an amplified ribosomal DNA restriction analysis (ARDRA), and phenotypically based on their colonial morphology, physiology, and nutritional requirements. Three different 16S rRNA gene-based genotypes were observed at a 100% similarity using the enzymes HinfI, MspI, and RsaI, and the phenotypic results also followed the ARDRA groupings. Selected strains, representing the different ARDRA groups, were analyzed by 16S rDNA sequencing, and members of the genera Halobaeillus, Virgibacillus, and Oceanobacillus were found. Two isolates showed low 16S rDNA sequence similarities with the closest related species of Halobacillus, indicating the presence of new species among the isolates. The majority of the strains isolated in this study seemed to belong to the species O. iheyensis and were compared using an AP-PCR to determine whether they had a clonal origin or not. Different patterns allowed the grouping of the strains according to Pearson's coefficient, and the resulting dendrogram revealed the formation of two main clusters, denoted as A and B. All the strains isolated from the soil were grouped into cluster A, whereas cluster B was exclusively composed of the strains associated with the B. portulacoides roots. This is the first report on the isolation and characterization of halotolerant spore-forming Gram-positive bacteria that coexist with B. portulacoides. As such, these new strains may be a potential source for the discovery of bioactive compounds with industrial value.

Microbial Community in the TPH-Contaminated Aquifer for Hot Air Sparging using Terminal-Restriction Fragment Length Polymorphism (유류오염대수층 고온공기분사공정시 제한효소다형성 미생물 군집)

  • Lee, Junho;Park, Kapsong
    • Journal of Korean Society on Water Environment
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    • v.24 no.1
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    • pp.19-29
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    • 2008
  • Hot air sparging is a groundwater remediation technique, in which organic contaminants volatilized into hot air from the saturated to vadose zone. In the laboratory diesel (10,000 mg TPH/kg) was spiked in contaminated saturated aquifer soil. The hot air ($34.9{\pm}2.7^{\circ}C$) was injected in intermittent (Q=1,500 mL/min, 10 minute injection and 10 minute idle) modes. We performed microcosm tests using the groundwater samples to assess TPH reductive remediation activity. For Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis of eubacterial communities in sludge of wastewater treatment plants and soil of experiment site, the 16S rDNA was amplified by Polymerase Chain Reaction (PCR) from the sludge and the soil. The obtained 16S rDNA fragments were digested with Msp I and separated by electrophoresis gel. We found various sequence types for hot air sparging experiment with sludge soil samples that were closely related to Bacillus (149 bp, Firmicutes), Methlobacterium (149 bp, Euryarchaeotes), Pseudomonas (492 bp, ${\gamma}$-Proteobacteria), etc., in the clone library. In this study we find that TPH-water was reduced to 78.9% of the initial value in this experiment aquifer. The results of the present study suggests that T-RFLP method may be applied as a useful tool for the monitoring in the TPH contaminated soil fate of microorganisms in natural microbial community.

Electrochemical Characteristics of Carbon/Carbon Hybrid Capacitor and Li-ion Battery/Hybrid Capacitor Combination (Carbon계 Hybrid Capacitor의 전기 화학적 기술 및 Li-ion Battery의 혼성 동력원 특성)

  • Lee, Sun-Young;Kim, Ick-Jun;Moon, Seong-In
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2005.07a
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    • pp.597-598
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    • 2005
  • Recently, the performance of portable electric equipment can often improved by a Li-ion battery assisted by a supercapacitor. A supercapacitor can provide high power density as well as a low resistance in the hybrid system. In this study, we have prepared, as the pluse power souce, a commercially supplied Li-ion battery with a capacity of 700mAh and AC resistivity of $60m\Omega$ at 1kHz and nonaqeous asymmetric hybrid capacitor composed of an activated carbon cathode and MCMB anode, and have examined the electrochemical characteristics of hybrid capacitor and the pulse performances of parallel connected battery/hybrid capacitor source. The nonaqueous asymmetric hybrid capacitor, the stacks of 10 pairs of the cathode, the porous separator and the anode electrode were housed in Al-laminated film cell. The hybrid capacitor, which was charged and discharged at a constant current at $0.25mA/cm^2$ between 3 and 4.3V, has exhibited the capacitance of 100F. And the equivalent series resistance was $32m\Omega$ at 1kHz. By combining a Li-ion battery and a hybrid capacitor, the pulse performance of battery can be improved 23% in run time under a pulse discharge of 7C-rate.

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ELISA detection of vivax malaria with recombinant multiple stage-specific antigens and its application to survey of residents in endemic areas

  • Kim, Sera;Ahn, Hye-Jin;Kim, Tong-Soo;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.41 no.4
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    • pp.203-207
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    • 2003
  • An ELISA was developed for the diagnosis of vivax malaria using multiple stage-specific recombinant antigens of Plasmodium vivax. The DNA from the whole blood of a malaria patient was used as template to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1). Each amplified DNA fragment was inserted into pQE30 plasmid to induce the expression of His-tagged protein in Escherichia coli (M15 strain) by IPTG. His-tagged proteins were purified by Ni-NTA metal-affinity chromatography and used as antigens for ELISA with patient sera that were confirmed previously by blood smear examinations. When applied to patient sera, 122 (80.3%) out of 152 vivax malaria cases reacted to at least one antigen, while no reactions were observed with 128 uninfected serum samples. We applied this ELISA to the screening of 3,262 civilian residents in endemic regions near the DMZ, which resulted in 236 positively detected (7.2%) cases. This method can be applied to serological diagnosis and mass screening in endemic regions, or can be used as a safety test for transfusion blood in endemic areas.

Functional Screening for Cell Death Suppressors and Development of Multiple Stress-Tolerant Plants

  • Moon Hae-Jeong;Baek Dong-Won;Lee Ji-Young;Nam Jae-Sung;Yun Dae-Jin
    • Journal of Plant Biotechnology
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    • v.5 no.3
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    • pp.143-148
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    • 2003
  • Bax, a mammalian pro-apoptotic member of the Bcl-2 family induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorhodamine123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MSP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to playa novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.