• Title/Summary/Keyword: Z-DNA

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Cloning and Sequencing of the Gene Involved in Morphological Change of Zoogloea ramigera 115SLR

  • Lee, Sam-Pin;Kim, Tae-Rahk;Sinskey, Anthony-John
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.161-168
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    • 2000
  • Plasmid pLEX3 isolated from the recombinant cosmid library of Zoogloea ramigera 115 was found to be responsible for the restoration of the rugose colony phenotype. To confirm the essential region responsible for the complementation, subclones were constructed from plasmid pLEX3 and transformed into mutant strain Z. ramigera 115SLR. The recombinant plasmids pLEX10 and pLEX11 were shown to complement the slime-forming property of Z. ramigera 115SLR. In a compositional analysis of the exopolysaccharides from Z. ramigera 115, Z. ramigera 115SLR, and Z. ramigera 115SLR harboring plasmid pLEX11, the exopolysaccharides showed a similar composition with glucose, galactose, and side chain groups. The complete nucleotide sequence of the 3.25kb genocim DNA insert in plasmid pLEX11 was determined and its analysis identified two open reading frames which could encode two proteins. The gene products derived form the two open reading frames were confirmed by and in vivo transcription using a T7-RNA polymerase. The ORF1 produced a 30 kDa protein, whereas the ORF2 was found responsible for the complementation of the morphological mutation and produced a 14 kDa protein. An in vivo gene expression of plasmid pTEX10 showed another open reading frame encoding a 50 kDa protein. The gene products form ORF1 and ORF2 are regarded as novel proteins which do not show any homology with other proteins.

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DNA Repair Capacity in Peripheral Blood Lymphocytes Predicts Efficacy of Platinum-based Chemotherapy in Patients with Gastric Cancer

  • Zhang, Yi-Yin;Gu, Kang-Sheng
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5507-5512
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    • 2013
  • Objective: To investigate the correlation between ERCC1 expression levels in tumor tissue and peripheral blood lymphocytes (PBL) from patients with gastric cancer and assess the relationship between PBL DNA repair rate (DRR) and the efficacy of platinum chemotherapy. Methods: A total of 53 patients with gastric cancer receiving surgery and 20 controls were studied. ERCC1 protein expression in tumour tissue and PBL were determined by immunohistochemical staining. The PBL DRRs of 47 advanced patients and 20 controls were estimated by comet assay. Results: The positive expression rates of ERCC1 were 67. 9%, 56. 6% and 10.0% in tumour tissues, PBLs of gastric cancer patients, and PBLs of the control group. PBL ERCC1 expression correlated with that in tissue (${\chi}^2$=15. 463, p=0.000). Pearson contingency coefficient=0.475). DRRs of cancer patients by tail length (TL) (Z=4. 662, p=0.000) and tail moment (TM) (Z=3. 827, p=0.000) were significantly lower than that of control group. When TL was applied as an indicator, the correlation between DRR and chemotherapy efficacy was significant (Spearman rank correlation r=0.327, p=0.032). Patients with low levels of DRR in PBL presented better short-term efficacy of chemotherapy than those with high levels of DRR. Conclusions: The ERCC1 expression in PBLs may indirectly reflect ERCC1 expression in gastric cancer tissues. Compared with non-cancer populations, patients with gastric cancer may have lower DNA repair capacity. DRR in PBL may predict the short-term efficacy of platinum-based chemotherapy for patients with advanced gastric cancer.

Cloning of SNAS-25 Gene from Rat Brain cDNA Library (Rat Brain cDNA Library로부터 SNAP-25 유전자의 클로닝)

  • Cho, Ae-Ri;Ji, Young-Mi;Yoo, Min;Lee, Soon-Chul;Yoo, Kwan-Hee
    • Biomedical Science Letters
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    • v.6 no.1
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    • pp.11-17
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    • 2000
  • SNAP-25 was first investigated as a neuron-specific protein preferentially expressed in CA3 pyramidal neurons of mouse hippocampus. It is a presynaptic plasma membrane protein in the nerve cell and plays an important role in the synaptic vesicle membrane docking and fusion pathway. We have recently isolated SNAP-25 cDNA from a rat brain cDNA library using a probe of Z2 cDNA. It consisted of 2,101 bp and an open reading frame (ORF) was identified between nucleotides (nt) 209 and 827. The AUG codon (nt 209∼211) was surrounded by CTACCATGG, which corresponded to the consensus sequence of ribosomal binding site. The ORF was terminated by TAA (nt 827∼829) to encode a polypeptide of 206 amino acid residues. The 3'-untranslated region contained two extensive stretches of repeated (CA)28 and (CA)19 at positions 925∼980 and 1645∼1682. It is noteworthy that cysteine residues were clustered in the span of amino acid residues 84∼991 : Cys-Gly-Leu-Cys-Val-Cys-Pro-Cys. Rat SNAP-25 showed 88% and 97% identity in nucleotide sequences to that of human and mouse, respectively. Amino acid sequence of rat SNAP-25 showed 100% identity to that of mouse and human SNAP-21.

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Analysis of the Molecular Mechanism of nlp Gene Involved in Transcriptional Regulation in Escherichia coli (대장균의 전사조절 유전자 nlp의 분자기구 해석)

  • 최용락;정수열;정정한;정영기
    • Microbiology and Biotechnology Letters
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    • v.21 no.3
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    • pp.229-238
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    • 1993
  • An nlp (Ner like protein) gene from E. coli was previously cloned and sequenced. Here we show that expression of the sugar metabolism related genes, lacZ, malQ and malP, increased 2.5-to 8.3-fold in the presence of a plasmid containing the nlp gene. This suggested that the nlp gene could induce maltose- and lactose-metabolism coordinately with crp*1 in the absence of cAMP. Using the nlp-lacZ fusion gene, it was possible to show the promoter of nlp was active in vivo.

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Zic3z Defines the Dorsal and Vegetal Neuroectoderm in the Zebrafish Embryonic Development

  • Lee, Kyu-Sun;Huh, Tae-Lin;Lee, Chang-Joong;Rhee, Myung-Chull
    • Animal cells and systems
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    • v.12 no.1
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    • pp.23-33
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    • 2008
  • The Zic family is a group of genes encoding zinc finger proteins that are highly expressed in the mammalian cerebellum. Zic genes are the vertebrate homologue of Drosophila pair-rule gene, odd-paired(opa), which plays important roles in the parasegmental subdivision as well as in the visceral mesoderm development of Drosophila embryos. Recent studies on human, mouse, frog, fish and ascidian Zic homologues support that Zic genes are involved in a variety of developmental processes, including neurogenesis, myogenesis, skeletal patterning, and left-right axis establishment. In an effort to explore possible functions of Zic proteins during vertebrate embryogenesis, we initially examined more detailed expression pattern of zebrafish homologue of zic3(zic3z). zic3z transcripts are detected in the neuroectoderm, neural plate, dorsal neural tube, and brain regions including eye field during early embryonic development. Marker DNA studies found that zic3z transcription is modulated by BMP, Wnt, and Nodal signals particularly in the dorsal and vegetal neuroectoderm at gastrula. Interfering with zic3z translation with zic3z-specific morpholino causes abnormal brain formation and expansion of the optic stalk cells. Retinal ganglion cells(RGCs) undergo abnormal neuronal differentiation. These findings suggest that zic3z defines the dorsal and vegetal neuroectoderm to specify brain formation and retinal neurogenesis during early embryonic development.

New ecological health assessment approaches of an urban stream using molecular and physiological level biomarkers and bioindicators

  • Kim, Ja-Hyun;Yeom, Dong-Hyuk;Kim, Joon-Ha;An, Kwang-Guk
    • Animal cells and systems
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    • v.16 no.4
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    • pp.329-336
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    • 2012
  • This study evaluated ecological health, using various biomarkers and bioindicators, of pale chub (Zacco platypus) as a sentinel species, in Daejeon Stream, South Korea, during AprilMay 2011. The biomarkers and bioindicators were compared among three sites of control: Reference ($C_z$), transition ($T_z$), and the urban zones ($U_z$); and the 7-Ethoxyresorufin-O-deethylase (EROD) activity, DNA damage, acetylcholinesterase (AChE) activity, and vitellogenin (VTG) concentrations were more significantly increased in the $U_z$ than in the $C_z$. Also, physiological markers such as condition factor, liver somatic index, visceral somatic index, and gonad somatic index were significantly increased in the $U_z$ than in the $C_z$. For the health assessments, three categorized parameters of blood chemistry, molecular biomarkers, and physiological bioindicators were standardized and calculated as a star-plot, representing values of Integrated Health Response (IHR). Values of IHR had more significant (P<0.05) increases in the $U_z$ than any other zones, indicating an impairment of ecological health by organic matter, nutrients (N, P), and toxic chemicals. This study is based on low levels of biological organization approach of molecular and physiological biomarkers and bioindicators, so further study of high-levels of biological organization approach such as community and population is required for overall range of health assessments. The approach of IHR values, however, may be useful in providing early warning of future impacts on ecological health.

In Vivo Transfer of Foreign DNA into Primordial Germ Cells (PGCs) of Chicken Embryos

  • Eguma, K.;Soh, T.;Hattori, M.;Fujihara, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.4
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    • pp.520-524
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    • 1999
  • The present experiments were designed to examine whether exogenous DNA injected into the germinal crescent region (GCR) of early stage of developing embryos, which is considered to be the main place from which PGCs originate, can be transferred to recipient chicken embryos. In this experiment, Miw Z (DNA) dissolved in the transfection reagent (TR: Boehringer, Germany) was introduced into the GCR of donor embryos at stage 3-5 or 9-11, followed by continued incubation until the stage 13-15 of embryonic development. The PGCs collected from the embryonic blood vessels were examined for the incorporation of the injected DNA into the PGCs by the methods of X-gal staining and PCR analysis. As the results, the foreign DNA was successfully incorporated into the PGCS, leading to their transfer to the gonadal tissues. The present results, therefore, suggest that the early stage (3-5 or 9-11) of chicken embryonic development would be more successful than stage 13-15 in transferring exogenous genes to the recipient embryos, leading to the possibility of producing transgenic chicken medianting the PGCS.

Phylogeographic Messages Encoded in the rDNA of the Commercial Mushroom Zhenghonggu@ From Fujian, China

  • Chen, Yu H.;Chen, Peng D.;Chen, Liu Y.;Ma, Li Z.
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.45-45
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    • 2014
  • Individualities of precious health mushroom called Zhenghonggu@ from respective protections scattered among all main mountains of Fujian China were collected and recognized locally, then compared with Russula griseocarnosa. Their internal transcribed spacer (ITS) region (ITS1, ITS2 and 5.8S rDNA) of the nuclear rDNA were amplified, AMOVA analyzed, nested clade analyzed and then compared with the ITS sequences of relative Russula species from other regions of China to confirm the taxonomic status of Zhenghonggu$^@$ and its population structure. Total 23 haplotypes from different protections of Fujian can be clustered into three clades similar to the three lineages of Dahongjun$^@$ from southeastern China reported by Li et al. The geographic distribution characteristic of these three phylogeny clades may be closely coupled with the vegetation regionalization and/or the differences of coenosium construction of Fagaceae that is the host of Russula griseocarnosa. The correlation of taxonomy, phylogeny and geographical distribution of Russula are discussed.

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Molecular Cloning and Characterization of Type 2 Metallothionein cDNA from Codonopsis lanceolata (S. et Z.) Trautv

  • In, Jun-Gyo;Lee, Bum-Soo;Yi, Tae-Hoo;Yu, Chang-Yeon;Yang, Deok-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.5
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    • pp.288-292
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    • 2005
  • A class I type 2 metallothionein (CMet2) cDNA from taproot of Codonopsis lanceolata was isolated and characterized. A CMet2 cDNA was 572 nucleotides long and had an open reading frame of 234 bp with a deduced amino acid sequence of 78 residues (pI = 4.99). The deduced amino acid sequence of CMet2 matched to the previously reported type 2 metallothionein-like protein genes and showed 74% identity with that of G. max (BAD18377) and C. arietinum (CAA65009). Expression of CMet2 by the RT-PCR was increased at 1 hr after cadmium and hydrogen peroxide treatment, respectively.

Regulatory Expression of DNA Repair Genes Involved in Adaptive Response (적응반응 관련 DNA 회복유전자의 발현조절에 관한 연구)

  • 최수영;이희원;박상대
    • Environmental Mutagens and Carcinogens
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    • v.10 no.1
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    • pp.1-10
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    • 1990
  • The regulation of DNA repair genes expression was investigated using fused genes, in which the promoter of repair genes was hybridized with the lacZ structural gene. The activities of beta-galactosidase expressed from the fused gense were highly increased when the host cells were exposed to methylating agents, such as methyl methansulfonate (MMS), N-methyl-N'-nitro-nitrosoguanidine (MNNG) and methyl nitrosourea (MNU). On the other hand, the enzyme activities from the fused genes were not induced when the cells were treated with ethylating or nonalkylating agents, such as ethyl methansulfonate (EMS), 4-nitroquinoline-1-oxide (4NQO), Bleomycin, and Benzo(a)pyrene (BP).

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