• Title/Summary/Keyword: KB-C2

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A Plausible Method for the Diagnosis of Genetic Disorders Using Full Length cDNA

  • Hur, Hyang-Suk;Lee, Young-Won;Park, Hyoung-Woo;Kim, Myoung-Hee
    • Biomedical Science Letters
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    • v.7 no.1
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    • pp.1-5
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    • 2001
  • A cDNA of coagulation Factor IX gene has been screened from the $\lambda$gt11 human fetal liver cDNA library, and used to construct a 2.8-kb full length cDNA after recombining with the N-terminal fragment from pTZ-FIX. Human genomic DNA was isolated, digested with the restriction endonucleases, TaqI, EcoRI, and HindIII, and Southern hybridization was performed using the full length factor IX cDNA as a probe. The hybridized bands generated by the restriction endonucleases were the followings: TaqI, 0.3, 1.0, 1.6, 1.8, 2.7, 3.7, and 5.3 kb bands; EcoRI, 1.8, 4.8, 4.9, 5.5, 6.8, and 12.6 kb bands; HindIII, 4.1, 4.4, 5.2, 5.8, 7.6, and 12.5 kb bands. When the Southern bands were physically mapped along the genome, about 50-kb continuous region harboring almost all of the genomic region of Factor Ⅸ gene was covered. These results suggest a possibility of using an exonal cDNA probe to diagnose abnormalities including large deletions, insertions, and rearrangements along the genome, if there is any.

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Cytogenetic and Molecular Genetic Studies on Duchenne Muscular Dystrophy (Duchenne Muscular Dystrophy에 관한 세포유전학 및 분자유전학적 연구)

  • Hong, Hea-Sook
    • Journal of Korean Biological Nursing Science
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    • v.7 no.1
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    • pp.29-46
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    • 2005
  • Purpose ; 본 연구는 X-염색체와 관련된 장애 중에서 가장 흔하고 심한 Duchenne Muscular Dystrophy(DMD)의 세포유전학 및 분자유전학적 특성을 설명하기 위해서 DMD에 영향을 받고 있는 두 가계의 13명을 대상으로 가계도 분석과 염색체 분석 및 DNA 분석을 하였다. Method ; DNA분석은 DNA probe을 이용한 Southern blotting method로써 RFLPs와 DMD유전자 부위의 exon소실 유무를 조사하여 아래와 같은 결과를 얻었다. Conclusion ; A 염색체 분석 : 말초혈액과 양수를 표본으로 High-Resolution GTG염색에서 A가계와 B가계의 염색체 분석에서 12명의 염색체는 정상 X-염색체였으나 B가계의 I-2(DMD여성)에서 46, x,-x,+t(2:x)(q 21.1 : p21.2)로 나타난다. B. DNA분석3 : 1) RFLPs의 분석 J66,XJ-1.1,754-11로써 B가계의 RELPs(Restriction Fragment Length Polymorphisms)에서 J66/Pst I은 1.7hb(E), 1.6kb(e)을 보여 주었고 XJ-1.1/Taq I은 3.6kb(F), 3.0kb(f), 754-11/EoR I은 4.2kb(G), 2.0kb(g)의 대립인자를 나타내었다. 이상의 결과를 바탕으로 영향을 받고 있는 남자 (II-2)의 haplotype는 보인자인 어머니의 한쪽 인자를 받았으며 어머니와 딸은 보인자이고 임산부의 태아는 남아였고 태아의 인자들은 그의 할아버지로부터 물려받아 DMD에 영향을 받지 않은 것으로 진단되었다. 2) DMD 유전자의 exon 소실에 대한 분석 cDNA probe 8과 cDNA probe 2b-3으로써 소실에 대한 진단은 영향을 받은 남자(II-2)는 cDNA probe 8에서 12, 7.3, 6.6, 4.2kb에 소실이 있고 cDNA 2b-3은 1.7kb에 소실에 나타났다.

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Rubiginone $B_2$, Isotetracenone Antibiotics which Reverses Multidrug-Resistance in KB Tumor Cells (KB 암세포에 효과있는 Streptomyces plicotosporus가 생산하는 항암증강물질 Rubiginone $B_2$ 에 관한 연구)

  • ;;Seto Haruo
    • Microbiology and Biotechnology Letters
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    • v.22 no.5
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    • pp.491-494
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    • 1994
  • Antibiotic HS-2 was purified from the culture broth of Streptomyces plicatosporus which was isolated from soil, by solvent extraction, silica gel column chromatography and gel filtration. Through the analysis of UV, $^{1}$H-NMR, $^{13}$C-NMR spectrum, HS-2 was identified as rubiginone B$_{2}$. It was confirmed that HS-2 enhanced the cytotoxicity of colchicine against multidrug-resistant tumor cells.

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Induction of Growth Inhibition by BCH in KB Human Oral Epidermoid Carcinoma Cells (구강 편평세포암종 KB세포에서 아미노산 수송억제제 BCH에 의한 세포성장 억제)

  • Yoon, Jung-Hoon;Kim, Youn-Bae;Kim, Do-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.5
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    • pp.758-763
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    • 2003
  • Amino acid transporters play an important role in supplying nutrients to normal and cancer cells for cell proliferation. System L is a major transport system responsible for the N $a^{+}$-independent, large neutral amino acids including several essential amino acids. L-type amino acid transporter 1 (LAT1), an isoform of system L amino acid transporter, is highly expressed presumably to support their continuous growth and proliferation in malignant tumors. 2-Aminobicyclo- (2,2,1) -heptane-2-carboxylic acid (BCH) is a model compound for study of amino acid transporter as a system L selective inhibitor. In the present study, we examined whether BCH induced growth inhibition in KB human oral squamous carcinoma cell line or not. The uptake of L-[$^{14}$ C]leucine by KB cells is inhibited by BCH in a concentration dependent manner with a Ι $C_{50}$ value of 75.3$\pm$6.2 ${\mu}{\textrm}{m}$ and a $K_{i}$ value of 98.7$\pm$ 4.1 ${\mu}{\textrm}{m}$. The growth of KB cells is inhibited by BCH in time dependent manner and concentration dependent manner with a Ι $C_{50}$ value of 11.1 $\pm$0.8 mM. In the DNA of KB cells treated with the various concentrations and various periods of BCH, the characteristic ladders associated with DNA fragmentation were not observed. These results suggest that BCH inhibits the growth of KB oral epidermoid carcinoma cells through the inhibition of transport of neutral amino acids into cells without DNA break down. This phenomenon will be a new rationale for anti-cancer therapy.y.

Characterization of RAD4 Homologous Gene from Coprinus cinereus (균류 Coprinus cinereus에서 DNA 회복에 관여하는 RAD4 유사유전자의 분리와 특성)

  • Choi, In-Soon
    • Journal of Life Science
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    • v.13 no.4
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    • pp.522-528
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    • 2003
  • The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the RAD4 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. In order to investigation whether the increase of transcripts by DNA damaging agent, transcripts levels were examined after treating the cells. The level of transcript did not increase by untraviolet light (UV). This result indicated that the RAD4 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the RAD4 homologous gene is essential for cell viability.

Effects of Sodding and Seeding Time and Rate of Seed Mixture on the Establishment of Cool-Season Turfgrasses (한지형 잔디의 조성시기, 초종 혼합 비율이 잔디 피복에 미치는 영향)

  • Shim Gyu Yul;Kim Chang Soo;Lee Seong Ho;Joo Young Kyoo
    • Asian Journal of Turfgrass Science
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    • v.18 no.4
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    • pp.179-191
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    • 2004
  • This study was conducted to find out the effect of sodding and seeding time and rate of seed mixtures on the establishment of cool-season turfgrasses by evaluating the turf coverage rates for two years. In fall planting, the required establishment period of full coverage($100\%$) was 1.5 months with a rolled turf sodding(Kentucky bluegrass $100\%$, Kentucky bluegrass $80\%$+perennial ryegrass $20\%$). The $100\%$ turf establishment was achieved in 7 months with Perennial ryegrass $100\%$, and 7.5 months by seeding with Kentucky bluegrass $100\%$(KB 100), Kentucky bluegrass $80\%$+perennial ryegrass $20\%$(KB80+PR20), Kentucky bluegrass $70\%$+perennial ryegrass $30\%$(KB70+PR30). In spring planting, the establishment periods far sod with KB 100 or KB80+PR20 were taken one month. However, in the case of seeding, the establishment periods were 3 months, 3.5 months, 3.5 months and 4 months with PR100, KB80+PR20, KB70+PR30, and KB 100, respectively Comparing the turf establishment vigor between fall and spring planting, the vigor was higher In spring planting than in fall planting in both sodding and . seeding. In the case of spring planting, the most proper time for turf establishment was tested on April, May, and June trials. The effect was significant in establishment vigor. The result showed highest on April planting. On May and June trials, establishment vigors were decreased gradually As the mixture rate of PR increased, ryegrass, establishment vigor was decreased with the rates. These results indicated that perennial ryegrass has relatively less tolerant to summer heat than Kentucky bluegrass. Number of shoots in 95 days after seeding was higher in KB100 by 16,600 per $m^2$ than in PR100 by 12,400 per $m^2$, while the lowest number showed in KB50+PR50 by 3,300 per $m^2$. Those in KB80:PR20, KB70:PR30 were 6,700 and 4,900 per $m^2$, respectively. The ratios of tillers according to mixture rates between Kentucky bluegrass and perennial ryegrass were KB80:PR20=87:13, KB70:PR30=78:22, and KB50:PR50=48:52. According to results in this study, Ideal seeding time might be spring (April) than in fall (September), and proper mixture rate was $80\%$ of Kentucky bluegrass with $20\%$ of perennial ryegrass.

Garnet-Orthopyroxene Geothermometer and Geological Applications (석류석-사방휘석 지질온도계와 지질학적 응용)

  • Lee, Han Yeang
    • Economic and Environmental Geology
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    • v.21 no.1
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    • pp.29-44
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    • 1988
  • Equilibrium relations between garnet and orthopyroxene have been investigated by reversal experiments in the range of 20-45Kb and $975-1400^{\circ}C$ in the $FeO-MgO-Al_2O_3-SiO_2$(FMAS) system. A mixture of PbO with about 55 mol per cent $PbF_2$ was used as a flux and proved very effective. The Fe-Mg exchange reaction seems to have little or no compositional dependence at these conditions. Combination of the experimental results with the garnet mixing model of Ganguly and Saxena(1984) yields the following geothermometric expression for the common natural assemblages that can be represented essentially within the system $FeO-MgO-CaO-MnO-Al_2O_3-SiO_2$. $$T^{\circ}C=(1971+11.91P(Kb)+1510(X_{Ca}+X_{Mn})^{Gt}/(lnK_D+0.96)-273$$.

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BCR (Brown Color Repressor) gene isolation related to mycelial browning of Lentinus edodes (표고균사 갈변과 관련된 BCR (Brown Color Repressor) 유전자 분리)

  • Kim, Young-Ho;Park, Soo-Cheol;Jhune, Chang-Sung;You, Chang-Hyun;Sung, Jae-Mo;Kong, Won-Sik
    • Journal of Mushroom
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    • v.10 no.3
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    • pp.120-128
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    • 2012
  • Recently sawdust cultivation of Shiitake mushroom (Lentinula edodes) is increasing. It is important to make mycelia to be brown on the substrate surface. This browned surface in sawdust cultivation plays an important role like as artificial bark of the oak log, which protects the other pests and suppresses water evaporation in the substrate. In order to isolate genes which related to brown color formation, differential display method was used. Two cDNA fragments obtained by DD-PCR were 1.2 and 1.6kb and these were expressed in white colored mycelia from L. edodes, but not brown colored mycelia. Partial sequencing of these cDNA fragments showed that the 1.6kb cDNA had 100% identity with the microsatellites gene from Dugenia polichroa. However, the other 1.2kb cDNA fragment had poly T tail on 3' region of partial open reading frame on 5' region. The new primer designed based on the sequence of 1.2kb cDNA was constructed. RT-PCR analysis using the newly designed 0.12kb cDNA specific primer showed that the gene was only expressed in white color mycelia, but not in brown color mycelia. Sequence analysis of 5' region of this 1.2kb cDNA revealed that this gene contained partial open reading frame consisted of 110 amino acid. Homology search using DNASIS database showed that this gene had high sequence homology of 66.7% in DNA level and 69.2 % in amino acid level with dTDP-glucose 4,6-dehydratases gene from Arabidopsis thaliata. The dTDP-glucose 4,6-dehydratases gene was known to be function to have tolerance with oxidation stress. These results strongly suggest that this gene isolated from white mycelia of L. edodes might have a function of repressor against mycelia browning. Therefore I designated this gene as BCR (Brown Color Repressor) gene.

Molecular Cloning of nifH, D from Frankia EuIK1 Strain, A Symbiont of Elaeagnus umbellata Root Nodules (보리수나무 뿌리혹 공생균주인 Frankia EuIK1의 nifH, D클로닝)

  • Kim, Ho-Bang;Kim, Chun-Ho;Song, Seun-Dal;An, Chung-Sun
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.258-263
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    • 1994
  • Genomic Southern hybridization of Frankia EuIKl strain, a nitrogen fixing symbiont of Elaeagnus umbellate root nodules, with nifH,D of K. pneumoniae as a probe, showed that 3.2 Kb and 5.5 Kb of BamHI fragments and 15 Kb PstI fragment were strongly hybridized with the probe, indicating nifH,D are located on these fragments. Using the same probe, one clone(pEuNIF) was isolated from the genomic library constructed into pWE15 cosmid vector by colony hybridization. The 3.2 Kb and 5.5 Kb BamHI fragments of this clone were hybridized with the same probe and this result corresponds to the genomic Southern hybridization data. However, using nifH of Frankia FaCl strain as a probe, only the 3.2 Kb BamHI fragment showed hybridization signal. Amino acid sequence deduced from nucleotide sequence of 3' terminus of the 3.2 Kb and 5' terminus of the 5.5 Kb fragments showed that the former was highly homologous with that of ArI3 nifD from 182nd to 240th amino acids, while the latter was from 241st to 282nd amino acids. These results show that nifH and partial nifD sequences are located on the 3.2 Kb fragment and residual sequences of nifH on the 5.5 Kb fragment which is contiguous to the 3.2 Kb fragment.

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Localization of MAK18 gene on chromosome VIII of saccharomyces cerevisiae (Saccharomyces cerevisiae의 염색체 VIII상의 MAK 18 유전자 국소화)

  • 윤순찬;이현숙;이창원
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.318-323
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    • 1988
  • MAK18 gene of Saccharomyces cerevisiae, needed for M1 replication, was mapped within 2cM of PET3 on chromosome VIII. From 38kb clone pRE66 carrying SPO11 and PET3, we have localized MAK18 gene whose insert is 2.8kb. MAK18 gene is Iocalized on about 9kb distance from PET3 and about 18kb distance from SPO11 on chromosome VIII.

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