• Title/Summary/Keyword: Northern hybridization

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Characterization of UV-Inducible Gene (UVI-180) in Schizosaccharomyces pombe (분열형 효모 Schizosaccharomyces pombe에서 자외선 유도유전자 UVI-180의 특성 연구)

  • Park, In-Soon
    • Environmental Analysis Health and Toxicology
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    • v.18 no.3
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    • pp.225-230
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    • 2003
  • 본 연구는 DNA 상해유도기작을 규명하기 위하여 하등 진핵생물인 분열형 효모 Schizosaccharomyces Pombe로부터 subtraction hybridization방법을 이용하여 자외선 유도 유전자인 UVI-180을 분리하고 그 유전자 구조와 발현양상을 조사하였다. UVI-180유전자의 발현양상을 Northern hybridization 방법으로 살펴본 결과 자외선(ultraviolet-light)조사 1시간 후에 최대의 발현 증가를 나타내었다. 반면 알킬화제인 MMS(methyl methanesulfonate)처리에 의해서는 전혀 발현이 증가되지 않았다. 이 결과 UVI-180유전자는 DNA상해에 따라 각기 다른 발현양상을 나타냄을 알 수 있었다. 유전자의 기능을 알기 위하여 null-mutant세포 주를 제조하여 그 특성을 살펴본 결과 이 유전자는 세포의 성장에 필수적인 유전자임을 알 수 있었다.

Isolation and Characterization of UV-Inducible Gene UV150 and UV200 in Eukaryotic Cells (진핵세포에서 DNA 상해에 반응하는 유전자 (UV150과 UV200) 기능연구 분리 및 특성 연구)

  • Choi In-Soon
    • Environmental Analysis Health and Toxicology
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    • v.21 no.1 s.52
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    • pp.21-26
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    • 2006
  • 본 연구는 DNA 상해유도기작을 규명하기 위하여 하등 진핵생물인 분열형 효모 Schizosaccharomyces pombe로부터 subtraction hybridization방법을 이용하여 자외선 유도 유전자인 UV150과 UV200을 분리하고 그 유전자 구조와 발현양상을 조사하였다. 분리한 유전자의 발현양상을 Northern hybridization 방법으로 살펴본 결과 자외선 조사 1시간 후부터 발현이 증가되었다. 또한 알킬화제인 Methyl Methanesulfonate (MMS) 처리에 의해서도 발현이 증가되었다. 이 결과 다른 UV-inducible유전자와는 다르게 분리한 UV150유전자는 UV에 UV200유전자는 MMS에 의하여 발현이 증가됨을 알 수 있었다. 유전자의 기능을 알기 위하여 URA4 유전자를 이용하여 null-mutant 세포주를 제조하여 그 특성을 살펴본 결과 분리한 UV150 유전자는 세포의 성장에 필수적인 유전자임을 알 수 있었다.

Taxonomic Strudy of the Combitid Fish, Cobitis Iutheri Rendahl and C. striata Ikeda (Cobitidae) from Korea (한국산 점줄종개(Cobitis lutheri) 와 줄종개(C.striata)의 분류학적 연구)

  • 김익수;이금영
    • Animal Systematics, Evolution and Diversity
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    • v.4 no.2
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    • pp.91-102
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    • 1988
  • Both Cobitis lutheri Rendahl and C.striata Ikeda previously regarded as the subspecies of C.taenia are revised here and raised to the species rank based on the distinct color pattern on their body sides in relation to the shpae of lamina circularis and suborbital spine, and distinct distributional patter. C. lutheri was similar to C. striata in chromosome number and karyotype, but chromosomal polymorphism as Robert sonian event was confirmed only in the population of C.lutheri studies. Both, C. kutheri and C..striata have disjunct ranges : the former in western Korea and east-northern China Mainland, the latter in the Smjin River of korea and west-southern Japan. hybridization between C. lutheri and C. striata by secondary contact appeared in the limited zone of the Dongjin River, Chllabuk-do province, korea, but the evidence for habitat segregation between them suggests the possibility that natural hybridization occurs between the two species and introgression results. We consider that the two species were produced as ecological equivalent species in the different branch stream of the Paleo-Hwangho River , The time of recession of sea level during the gracial period.

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Differentially Expressed Genes Related to Cold-resistance in Barley (Hordeum vulgare L. cv. Nagaoka)

  • Chun, Jong Un;Park, Jeong-Seon;Bae, Chang-Hyu;Shin, Jeong-Sheop
    • Korean Journal of Breeding Science
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    • v.41 no.2
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    • pp.73-83
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    • 2009
  • To investigate genes related to vernalization and cold- resistance in barley (Hordeum vulgare L. cv. Nagaoka), differentially expressed genes were identified from cold-resistant barley leaves with suppression subtractive hybridization (SSH) and Northern blot analyses. The nucleotide and the deduced amino acid sequences of the putative gene products were compared. The bvrn-7 showed high homology(84%) with gene related to vernalization, and the bvrn-3, bvrn-12, bvrn-28, bvrn-29 and bvrn-36 related to cold-resistant genes had high identity of 88~98% with low temperature-induced genes. The results indicate that the 6 genes were closely related to vernalization and cold-resistance during low temperature treatment.

Expression and Regulatory Analysis of Sporulation Gene (spo 5) in Schizosaccharomyces pombe (Schizosaccharomyces pombe 포자형성유전자 (spo 5)의 발현조절기구의 해석)

  • KIM Dong-Ju;SHIMODA Chikasi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.1
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    • pp.46-54
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    • 1997
  • Sporulation in the fission yeast Schizosaccharomyces pombe has been regarded as an important model of cellular development and differentiation. S. pombe cells proliferate by mitosis and binary fission on growth medium. Deprivation of nutrients especially nitrogen sources, causes the cessation of mitosis and initiates sexual reproduction by malting between two sexually compatible cell types. Meiosis is then followed in a diploid cell in the absence of nitrogen source. DNA fragment complemented with the mutations of sporulation gene was isolated from the S. pombe gene library constructed in the vector, pDB 248' and designated as pDB (spo 5)1. We futher analyzed six recombinant plasmids, pDB (spo 5)2, pDB(spo 5)3, pDB(spo 5)4, pDB(spo 5)5, pDB(spo 5)6, pDB(spo 5)7, and found each plasmids is able to rescue the spo 5-2, spo 5-3, spo 5-4, spo 5-5, spo 5-6, spo 5-7, mutations, respectively. Mapping of the integrated plasmid into the homologous site of the S. pombe chromosomes demonstrated that pDB (spo 5)1, and pDB (spo 5)R1 contained the spo 5 gene. Transcipts of spo 5 gene were analyzed by Northern hybridization. Two transcripts of 3.2 kb and 25 kb were detected with 5 kb Hind III fragment containing a part of the spo 5 gene as a probe. The small mRNA (2.5 kb) appeared only when a wild-type strain was cultured in the absence of nitrogen source in which condition the large mRNA (3.2 kb) was produced constitutively. Appearance of a 2.5 kb spo 5-mRNA depends upon the function of the mei1, mei2 and mei3 genes.

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Fusion Between TMPRSS2 and ETS Family Members (ERG, ETV1, ETV4) in Prostate Cancers from Northern China

  • Wang, Jian-Jiang;Liu, Yue-Xin;Wang, Wei;Yan, Wei;Zheng, Yu-Peng;Qiao, Lu-Dong;Liu, Dan;Chen, Shan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.4935-4938
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    • 2012
  • In this study we evaluated the frequency of fusion between TMPRSS2 and ETS family members (ERG, ETV1, ETV4) in prostate cancers in patients from northern China in order to explore differences in fusion rates among regions in northern and southern China, other parts of Asia, Europe, and North America. We examined 100 prostate cancer patients, diagnosed by means of prostate biopsy; fluorescence in situ hybridization (FISH) was used to detect the expression of TMPRSS2, ERG, ETV1 and ETV4 in cancer tissue. Differences in gene fusion rates among different ethnics groups were also analyzed. Of the 100 prostate cancer patients, 55 (55%) had the fusion gene. Among the patients with the fusion gene, 46 (83.6%) patients had the TMPRSS2:ERG fusion product, 8 (14.8%) patients had TMPRSS2:ETV1 fusion, 1 (1.6%) patient had TMPRSS2:ETV4.

Expression of Multidrug Resistance-associated Protein (MRP), c-myc and c-fos in L1210 Cells (L1210 암세포에서 Multidrug Resistance-associated Protein (MRP), c-myc 및 c-fos 유전자의 발현양상)

  • Kim, Seong-Yong
    • Journal of Yeungnam Medical Science
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    • v.14 no.1
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    • pp.67-76
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    • 1997
  • The occurrence of multidrug resistance (MDR) is one of the main obstacles in the successful chemotherapeutic treatment of cancer. In this study The gene expressions of multidrug resistance-associated protein (MRP), c-myc and c-fos were investigated in L1210 cells. Adriamycin- or vincristine-resistant L1210 cells, L1210AdR or L1210VcR, respectively, has been identified to overexpression of mdr1 gene. The expression leve of MRP gene in L1210AdR and L1210Cis was more decreased than that in L1210 cells. The c-myc and c-fos genes were expressed both in L1210 and resistant sublines. In L1210AdR, the expressions level of c-myc and c-fos genes were decreased than in L1210. However, in L1210VcR and L1210Cis, c-myc and c-fosgene expressionwere rather increased than L1210. These results suggested that MRP does not contribute in resistance of drug-resistant L1210 cells and there is no relations between MRP and mdr1 gene expression. The expression of c-myc and c-fos gene may be changed during transformation of L1210 to drug-resistant sublines.

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Expression of protein kinase C in the spinal cords of rats with autoimmune encephalomyelitis (뇌염모델에서 Protein Kinase C의 발현에 관한 연구)

  • Shin, Tae-Kyun;Kim, Hyung-Min;Tanuma, Naoyuki;Matsumoto, Yoh
    • Korean Journal of Veterinary Pathology
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    • v.1 no.1
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    • pp.26-32
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    • 1997
  • Protein kinase C an enzyme of signal transduction has been known to regulate cell proliferation activation as well as apoptosis in some cancer cell lines. To explore the role of PKC in the course of cell mediated autoimmune disease such as experimental autoimmune encephalomyelitis (EAE) EAE was induced in Lewis rats(6-8 weeks old) with immunization of myelin basic protein supplemented with complete Freund's adjuvants and affected spinal cords were sampled at days 13 postimmunization(PI) as peak stage of EAE and at days 21 PI as recovery stage. The spinal cords with EAE were subjected to Northern blot analysis and insitu hybridization of PKC delta which is one of prominant isotypes of PKC in the haematopoietic cells. Northern blot analysis showed that levels of PKS delta mRNA in the spinal cords of rats withEAE was significantly increased at days 13 PI in which inflammatory cells including T cells and macrophages in the EAE lesions appeared. however the stage. By in situ hybridization signals of PKC delta in EAE lesions was intensely expressed on the delta is also expressed on some brain cells in normal rat central nervous system This finding suggests that PKC plays an important role on either activation of inflammatory cells including encephalitogenic T cells and macrophages or apoptotic elimination of some inflammatory cells depending on the stge of EAE.

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EFFECTS OF SUBSTANCE P ON COLLAGEN PRODUCTION IN HUMAN PERIODONTAL LIGAMENT CELLS (치주인대 세포의 교원질 생성에 대한 Substance P의 효과)

  • CHUN, Jun-Yeung;Choi, Je-Yong;Kyung, Hee-Moon;Sung, Jae-Hyun
    • The korean journal of orthodontics
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    • v.26 no.1 s.54
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    • pp.83-94
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    • 1996
  • Substance P is one of the neuropeptide which presents highly in tension site of periodontal ligament during the orthodontic tooth movement. It has bnn also hon as one of the neuropeptides which cause neurogenic inflammation in various tissues and organs. However, there is no report about the effect of substance P on major extracellular matrix protein, collagen production. The purpose of this study was to evaluate the collagen production by substance P in human periodontal ligament cell. The collagenase-digestion method was used to evaluate collagen production and also used Northern blot hybridization for the evaluation of collagen mRNA level. This study also Included in terms of prostanglandins and gelatinase production with respect to collagen production. For the collagen degradation, zymography was used to estimate denatured collagen degradation. Dose-dependent effect of substance P on noncollagen protein, collagen, and percent collagen was that substance P increased noncollagen protein synthesis, but decreased collagen sytnsis. So the percent collagen, which determined by relative collagen production against total protein production, w3s decreased from $7\%\;to\;3.6\%$. This inhibitory effect of substance P on collagen production was disappeared when cells were treated concomitantly with indomethacin. It means that substance P-induced inhibitory effect on collagen production was due at least in part to the production of prostaglandins. To evaluate whether substance P-induced inhibitory effect on collagen production is correspond to the steady-state levels of procollagen mRNA, Northern blot hybridization was performed and it showed that substance P has no effect on the steady-slate level of ${\alpha}1(I)$ procollagen mRNA. It means that the inhibitory effect of substance P on collagen production was due to the change of a certain mechanism after posttranscription. In this context, gelatinase production by substance P in periodontal ligament cells was evaluated by zymography. Zymogram showed that substance P has no effect on gelatinase production in periodontal ligament cells. To explore wheter substance P-induced inhibitory effect on collagen production is selevtive in periodontal ligament cells or not, MC3T3-E1 cells which originated from mouse calvaria was used. It showed that substance P has no effect on collagen production in MC3T3-E1 cells. Taken together, substance P inhibits collagen production in human periodontal ligament cells. This effect was not due to the change of the steady-state level of procollagen mRNA and gelatinase production, but due at least in part to the change of prostaglandins production.

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Construction of cDNA Library from Posterior Silk Gland (PSG) of Korean Oak Silkmoth, Antheraea yamamai and Molecular Cloning of Fibroin Heavy Chain Gene(FHC)

  • Lee, Jin-Sung;Kim, Soon-Jung;Kim, Ki-Hwan;Park, Young-Min;Suh, Dong-Sang
    • Journal of Life Science
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    • v.10 no.1
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    • pp.10-13
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    • 2000
  • To develope the genetic source of oak wild silkworm, Antheraea yamamai, the cDNA library was constructed with poly A+ mRNA isolated from posterial silk gland of fifth instar larvae. Titer of the cDNA library was about 5.1$\times$105 pfu in total. We presumed that the titer covered almost all transcripts existed in Antherea yamamai. From cDNA library of Antheraea yamamai, fibroin heavy chain gene, which is specifically expressed from posterial silk gland of Antheraea yamamai, was screened using oligonuclotide probe specific to alanine rich motif of fibrin heavy chain gene of Antheraea pernyi. As a result, fibroin clones isolated from 5$\times$104 plaques showed the highest homolgy (95%) with that of Antherea pernyi in nucleotide of Anthereaea yamamai and Bombyx mori shows that there is no homologous sequence in the 3+ partial 채야후 region Genomic southern hybridization suggested that one copy is present. Northern hybridization showed that fibroin transcript was approximateely 9 kb in length.