• 제목/요약/키워드: Cy5

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Construction of Complementary DNA Library and cDNA Cloning for Cy Strain of Odontoglossum Ringspot Virus Genomic RNA (오돈토글로썸 윤문 바이러스 Cy계통 게놈 RNA의 cDNA 구축 및 유전자 크로닝)

  • 류기현;박원목
    • Korean Journal Plant Pathology
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    • v.10 no.3
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    • pp.228-234
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    • 1994
  • Genomic RNA was extracted from Cy strain of odontoglossum ringspot tobamovirus (ORSV-Cy) isolated from infected leaves of tobacco cv. Samsun. Size of the genomic RNA was about 6.6 kb in length. The genomic RNA was fractionated using Sephadex G-50 column chromatography into 2 fractions. They were polyadenylated at their 3'-end using E. coli poly(A) polymerase. Polyadenylated viral RNA was recovered by oligo (dT) primer adapter containing NotI restriction site and Moloney murine leukemia virus SuperScript reverse transcriptase (RNase H-). Second-strand cDNA was synthesized by using E. coli DNA ligase, E. coli DNA polymerase I and E. coli RNase H. Recombinant plasmids containing cDNAs for ORSV-Cy RNA ranged from about 800 bp to 3,000 bp. Among the selected 238 recombinants, pORCY-124 clone was the largest one covering 3'-terminal half of the viral RNA. This clone contained two restriction sites for EcoRI and XbaI and one site for AccI, AvaI, BglII, BstXI, HindIII, PstI, and TthIII 1. respectively. The clone contained partial viral replicase, a full-length movement protein and a complete coat protein genes followed by a 3' untranslated region of 414 nucleotides based on restriction mapping and nucleotide sequencing analyses. Clones pORCY-028, -068, -072, -187 and -224 were overlapped with the pORCY-124. Clones pORCY-014 and -095 covered 5' half upstream from the middle region of the viral RNA, which was estimated based on restriction mapping and partial sequence analysis. Constructed cDNA library covered more than 90% of the viral genome.

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Studies on a New Wing Mutant (Surf Wings; Srf) of Drosophila melanogaster Extracted from a Wild Population (자연집단에서 초파리(Drosophila melanogaster)의 신돌연변이체(Sufr Wings;S f)에 관한 연구)

  • 姜永善;朴殷浩
    • The Korean Journal of Zoology
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    • v.14 no.2
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    • pp.75-84
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    • 1971
  • Genetic analysis of a new wing mutant, Surf wings (Srf), was performed. Mutant flies were extracted from a wild population of Drosophila melanogaster at the vicinity of Atomic Energy Research Institute, Seoul in August-September 1969. 1. The distal half of wings of heterozygotes (Srf/+) turned upwards about 40 degree from body axis, but flying ability was not disturbed. They overlap Cy in low frequency when they are grown below $22^{\circ}C$. This resembles with Si or j series, but wing margins are not rolled and diversed. Postscutellars are erected slightly, but they, in general, are not crossed. Any other external pleiotropic effects were not observed. 2. Penetrance and expressivity of both sexes are not complete. Their extents are variable with sex and temperature. These characters show maximum tendencies when the development is made at $22^{\circ}C$ (female: P = 0.996, E = 0.932, male:P = 0.961, E = 0.698). 3. The preliminary locus of Srf was determined to be 66.8 on the right arm of second chromosome by using recessive maker gene cn bw. 4. The homozygous flies(Srf/Srf) have shown perfect lethality. The heterozygotes (Srf/+), on the other hand, have shown to be viable and fertile. Srf chromosomes are kept in a balanced lethal system with Pm chromosomes which are associated with inversions. Hence, it is partially reasonable to suppose that Srf may persist in a natural population by the same mechanism. 5. Allelism test with Cy was also conducted. The fact that combination with Cy in the trans-phase (+ Srf/Cy +) is viable in contrast to the lethality of Srf/Srf and Pm/Pm indicates that Srf and Cy are not functionally allelic.

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Emulsifying and Gelling Properties of Pork Myofibrillar Protein as Affected by Various NaCl Levels and pH Values

  • Jang, Ho-Sik;Chin, Koo-Bok
    • Food Science of Animal Resources
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    • v.31 no.5
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    • pp.727-730
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    • 2011
  • The effects of various NaCl levels (0, 0.3, and 0.6 M) and pH values (pH 5.0, 5.5, 6.0, and 6.5) on the emulsifying and gelling properties of pork myofibrillar protein (MP) were assessed. The emulsion stability index (ESI), emulsifying activity index (EAI), and creaming index were measured at a 1:20 ratio of MP to corn oil. The EAI and ESI of pork MP showed maximum values at pHs 6.0 and 6.5 and at 0.3 M NaCl, resulting in better emulsion properties. Additionally, the cooking yield (CY) and gel strength (GS) of emulsified MP gel were measured at an MP: corn oil ratio of 1: 2; GS increased with increasing levels of salt. At 0.6 M NaCl, GS decreased with decreasing pH from 5.5 to 6.5. GS and gelling properties were optimal at pH 5.5 in 0.6 M salt. The highest CY was observed at 0.6 M NaCl, regardless of the pH value. However, increasing pH increased CY at salt levels of 0 and 0.3 M. These results indicate that NaCl and pH profoundly affected the emulsified MP system. Future work will be conducted on the rheological properties of the pork emulsified system as affected by adding non-meat protein.

Effect of missing values in detecting differentially expressed genes in a cDNA microarray experiment

  • Kim, Byung-Soo;Rha, Sun-Young
    • Bioinformatics and Biosystems
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    • v.1 no.1
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    • pp.67-72
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    • 2006
  • The aim of this paper is to discuss the effect of missing values in detecting differentially expressed genes in a cDNA microarray experiment in the context of a one sample problem. We conducted a cDNA micro array experiment to detect differentially expressed genes for the metastasis of colorectal cancer based on twenty patients who underwent liver resection due to liver metastasis from colorectal cancer. Total RNAs from metastatic liver tumor and adjacent normal liver tissue from a single patient were labeled with cy5 and cy3, respectively, and competitively hybridized to a cDNA microarray with 7775 human genes. We used $M=log_2(R/G)$ for the signal evaluation, where Rand G denoted the fluorescent intensities of Cy5 and Cy3 dyes, respectively. The statistical problem comprises a one sample test of testing E(M)=0 for each gene and involves multiple tests. The twenty cDNA microarray data would comprise a matrix of dimension 7775 by 20, if there were no missing values. However, missing values occur for various reasons. For each gene, the no missing proportion (NMP) was defined to be the proportion of non-missing values out of twenty. In detecting differentially expressed (DE) genes, we used the genes whose NMP is greater than or equal to 0.4 and then sequentially increased NMP by 0.1 for investigating its effect on the detection of DE genes. For each fixed NMP, we imputed the missing values with K-nearest neighbor method (K=10) and applied the nonparametric t-test of Dudoit et al. (2002), SAM by Tusher et al. (2001) and empirical Bayes procedure by $L\ddot{o}nnstedt$ and Speed (2002) to find out the effect of missing values in the final outcome. These three procedures yielded substantially agreeable result in detecting DE genes. Of these three procedures we used SAM for exploring the acceptable NMP level. The result showed that the optimum no missing proportion (NMP) found in this data set turned out to be 80%. It is more desirable to find the optimum level of NMP for each data set by applying the method described in this note, when the plot of (NMP, Number of overlapping genes) shows a turning point.

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Comparison of Hybridization Behavior between Double and Single Strand of Targets and the Application of Asymmetric PCR Targets in cDNA Microarray

  • Wei, Qing;Liu, Sanzhen;Huang, Jianfeng;Mao, Xueying;Chu, Xiaohui;Wang, Yu;Qiu, Minyan;Mao, Yumin;Xie, Yi;Li, Yao
    • BMB Reports
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    • v.37 no.4
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    • pp.439-444
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    • 2004
  • Double stranded targets on the cDNA microarray contain representatives of both the coding and noncoding strands, which will introduce hybridization competition with probes. Here, the effect of double and single strands of targets on the signal intensity and the ratios of Cy5/Cy3 within the same slide were compared. The results show that single stranded targets can increase the hybridization efficiency without changing the Cy5/Cy3 ratio. Based on these results, a new strategy was established by generating cDNA targets with asymmetric PCR, instead of conventional PCR, to increase the sensitivity of the cDNA microarray. Furthermore, the feasibility of this approach was validated. The results indicate that the cDNA microarray system based on asymmetric PCR is more sensitive, with no decrease in the reliability and reproducibility as compared with that based on conventional symmetric PCR.

Effects of Electrical Conductivity on the Soil Microbial Community in a Controled Horticultural Land for Strawberry Cultivation (시설딸기재배지 토양에서 염류농도가 미생물 생태에 미치는 영향)

  • Lee, Young-Han;Ahn, Byung-Koo;Sonn, Yeon-Kyu
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.5
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    • pp.830-835
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    • 2011
  • Total soil microbial activities have great impact to soil management for organic farming. This study was evaluated in the soil microbial community by fatty acid methyl ester (FAME) in a controlled horticultural field for strawberry organic farm. Experimental plots were prepared with a high level of soil electrical conductivity (EC) and a optimum level of soil EC. Soil microbial biomasses and communities of total bacteria, Gram-negative bacteria, Gram-positive bacteria, actinomycetes, fungi, and arbuscular mycorrhizal fungi in the high level of soil EC were significantly larger than those in the optimum level of soil EC. Lower ratios of cy17:0 to 16:$1{\omega}7c$ and cy19:0 to 18:$1{\omega}7c$ were found in the optimum level of soil EC than those in the high level of soil EC, indicating that microbial stress decreased.

Electrochemical Properties of Molydenum (V) Complexes with Multidentate Ligands Containing Nitrogen or / and Oxygen Donor Atom (질소, 산소 주게원자 리간드를 가진 몰리브덴 (V) 착물의 전기화학적인 성질)

  • Sang Oh OH;Soo Gyun ROH
    • Journal of the Korean Chemical Society
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    • v.33 no.5
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    • pp.484-495
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    • 1989
  • Reactions of $(Et_4N)_2[MoOCl_5]$ with multidentate ligands containing nitrogen or/and oxygen donor atom (EDTA, DTPA, IDA, CyDTA, OX) produce a series of binuclear molybdate (V) complexes. The prepared Mo (V) complexes has been identified by Elemental Analysis, Infrared Spectra, Proton Magnetic Resonance Spectra, and Electronic Spectra. The electrochemical reduction mechanism has been studied by Cyclic voltammetry, Controlled Potential Coulometry, and Spectrophotometry in pH 3.571-10.375 acetate, borate, phosphate/sodium hydroxide, phosphate, ammonium/ammonia buffers. The cyclic voltammogram of the Mo-EDTA, DTPA, IDA, CyDTA complexes at pH < ca. 6.00 have shown two reduction waves. The first reduction wave shows two electron process and the second reduction wave shows two electron process. The cyclic voltammogram of the Mo-EDTA, DTPA, IDA, CyDTA complexes at pH < ca. 8.00 has shown one reduction wave. This reduction wave show four electron process. The cyclic voltammogram of the Mo-OX complex at pH < ca. 7.2 has shown one reduction wave. This reduction wave show four electron process.

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Effect of Cysteine and Glutathione on In Vitro Maturation of Porcine Follicular Oocytes (Cysteine 및 Glutathione 이 돼지난포란의 체외성숙에 미치는 영향)

  • 신성진;한만희;이규승
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.385-393
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    • 1998
  • This study was conducted to investigate the effect of cysteine (CySH) and glutathione (GSH) on in vitro maturation of porcine follicular oocytes. The results obtained were summarized as follows: 1. When the immature oocytes were cultured at 0, 0.04, 0.14, 0.6 and 1.2 mM of cysteine (CySH) for 36h, the germinal vesicle breakdown (GVBD) rates were 90.8, 89.9, 90.5, 92.0 and 91.3%, respectively, and the maturation rates of the oocytes with metaphase-II were 56.1, 50.7, 41.9, 49.0 and 61.5%, respectively. Especially, the maturation rates of 0.14 and 0.6 mM treated groups were significantly lower than those of control (non-treated) group (P<0.05). After 44h of culture in the same treatments of CySH, the GVBD rates of porcine immature oocytes were 90.0, 91.8, 89.8, 90.5 and 89.6%, respectively, and the maturation rates were 80.2, 76.3, 69.4, 66.7 and 72.6%, respectively. Especially, the maturation rates of 0.14 and 0.6 mM treated groups were significantly lower than those of control group (P<0.05). 2. When the immature oocytes were cultured at 0, 0.05, 0.1, 0.5 and 1.0 mM of glutathione (GSH) for 36h, the GVBD rates of porcine immature oocytes were 91.0, 90.9, 89.5, 92.0 and 91.1%, respectively, and the maturation rates were 59.0, 48.5, 47.8, 38.6 and 37.5%, respectively. All treated groups of GSH showed lower maturation rates than the control group (P<0.05). After 44h of culture in the same treatments of GSH, the GVBD rates of porcine immature oocytes were 91.8, 94.1, 89.1, 91.3 and 91.1%, respectively, and the maturation rates were 84.6, 57.1, 69.6, 71.3 and 64.3% respectively. All treated groups of GSH showed lower maturation rates than the control group (P<0.05).

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Early and Late Effect of External Abdominal Irradiation on Small Intestine of Mice at Various Total Dose and Intervals - Histopathologc Aspects of Damage and Recovery - (복부 방사선 조사에서 총 선량 및 회복기간에 따른 마우스 소장의 급만성 효과 -손상 및 회복의 병리조직학적 변화-)

  • Kim, Myung-Se;Choi, Won-Hee
    • Radiation Oncology Journal
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    • v.5 no.1
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    • pp.1-11
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    • 1987
  • Total 390 mice were irradiated with $2{\times}3cm$ abdominal field and $200{\times}5/wk$ regimen with orthovoltage x-ray machine. All animals were divided into 2 large groups, damage and recovery, and histopathologic changes were analyzed at various total dose level (1,000 cCy-5,000 cCy) and intervals (1 week-15 weeks). Almost acute changes such as decreased mitotic activity, villi shortening and focal erosion of mucosa recovered within 1-2 weeks in 1,000-3,000 cGy irradiated group but mild changes persisted in 4,000 and 5,000 cCy irradiated groups. Chronic delayed changes such as thickening of vessel wall with focal thrombosis, submucosal fibrosis, mucosal atrophy and chronic ulcer were observed from 2-3 week specimens in 4,000 and 5,000 cGy groups, These late changes recovered slightly, as increasing time intervals after irradiation but mild, persistant changes were observed throught all follow up period. These data suggest hat even 4,000 cCy is not completely safe for possible permanent damage and judicious modificiation of total dose, dose rate, fraction size, and field size should be considered for better results.

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