• 제목/요약/키워드: lacI

검색결과 121건 처리시간 0.025초

유전자수준에서 돌연변이 유발기전을 밝히는 Transgenic Mutagenesis Assay (Transgenic Mutagenesis Assay to Elucidaate the Mechanism of Mutation at Gene Level)

  • 류재천;윤지윤;조경혜;장일무
    • 한국환경성돌연변이발암원학회지
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    • 제18권1호
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    • pp.15-21
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    • 1998
  • Transgenic animal and cell line models which are recently developed and used in toxicology fields combined with molecular biological technique, are powerful tools to study the mechanism of mutation in vivo and in vitro, respectively. Transgenic models, which have exogenous DNA incorporated into their genome, carry recoverable shuttle vector containing reporter genes to assess endogenous effects or alteration in specific genes related to disease processes. The lac I and lac Z gnee most widely used as a mutational target in transgenic systems. The assay is performed by treatment with putative mutagenic agents, isolation of genomic DNA from cells or tissues, exposure the isolated DNA to in vitro packaging extract, plating and sequencing. The results from these processes provide not only mutant frequency as quantitative evaluation but also mutational spectrum as qualitative evaluation of various agents. Therefore we introduce and review the principle, detailed procedure and application of transgenic mutagenesis assay system in toxicology fields especially in mutagenesis and carcinogenesis.

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유전자 재조합 대장균을 사용한 Alpha-interferon의 생산과 분비: 제2부. 재조합 균주의 생장특성 (Extracellular Production of Alpha-Interferon by Recombinant Escherichia coli: Part II. The Growth Behavior of the Recombinant Cells)

  • 노갑수;최차용
    • KSBB Journal
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    • 제5권3호
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    • pp.195-200
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    • 1990
  • 대장균의 lipoprotein promoter, lactose promotor 및 operator 와 lipoprotein의 signal sequence 를 가지는 vector에alpha-IFN 유전자가 cloning된 plasmid pIF-Ill-B를 여러종류의 대장균 숙주 세포에 형칠전환하여 alpha-IFN 의 생산성, 생장특성을 조사하였다. 또한 plasmid 자체와 cloning된 alpha-IFN 유전자의 발현유도가 세포생장에 미치는 영 향을 조사하였다.

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Differential Diagnosis of Brain Diseases Using In Vivo Proton Magnetic Resonance Spectroscopy at 3 Tesla: A Preliminary Study

  • Shen, Yu-Lan;Kang, Heoung-Keun;Kim, Tae-Hoon;Sundaram, Thirunavukkarasu;Kim, Hyeong-Jung;Jeong, Gwang-Woo
    • 한국자기공명학회논문지
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    • 제13권2호
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    • pp.64-83
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    • 2009
  • The purpose of this study was to evaluate the usefulness of in vivo 3T $^1H$ MRS with short TE for prescreening various brain diseases. Together with ten normal volunteers, 12 brain tumor patients(2 lymphomas, 5 malignant gliomas) and 5(benign meningiomas) and 10 brain ischemic disease patients(6 acute and 4 subacute infarctions) participated. Lymphomas showed increased intensities of Cho and Lac. Likewise, gliomas showed increased Cho and Lac, but with decreased NAA and ${\beta}\;{\gamma}$-Glx; in higher grade of gliomas, Lac, Cho, mI and Lip predominantly increased with decrease of NAA. Benign meningiomas showed increased Cho, Lac and ${\beta}\;{\gamma}$-Glx; with decreased of NAA. The alanine peak at 1.47 ppm is a neuronal marker for meningiomas. Infarctions showed increased Lac and Lip and decreased NAA, ${\alpha}$-Glx and ${\beta}\;{\gamma}$-Glx where Lac increased with decreased of ${\alpha}$-Glx in acute, and Cho, Lac and Lip increased with decrease of NAA in subacute. Elevated Lac and decreased NAA levels were more aggravated in subacute. Clinical application of the $^1H$ MRS with short TE at 3T is able to povide valuable spectral information for prescreening various brain diseases by monitoring the changes of disease-specific cerebral metabolite concentrations in vivo, and consequently, it can be applicable to assessment of differential diagnosis and malignancy as well.

젖산균과 온도가 김치발효에 미치는 영향(I) (Effect of Lactic Acid Bacteria and Temperature on Kimchi Fermentation (I))

  • 조영;이혜수
    • 한국식품조리과학회지
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    • 제7권1호
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    • pp.15-25
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    • 1991
  • 김치 발효 중에 주로 나타나는 Lac. plantarum, Leu. mesenteroides, Ped. acidilactici, Lac. brevis 등의 젖산균들이 김치 발효에 미치는 영향을 규명하고자, 이 젖산균들을 순수배양하여 염도 2.31%의 김치즙에 접종한 후 $30^{\circ}C$, $21^{\circ}C$, $14^{\circ}C$, $7^{\circ}C$에서 발효시키면서 pH및 총산도의 변화, 휘발성 유기산과 비휘발성 유기산의 변화를 관찰하였다. Control group과 Leu. mesenteroides를 접종한 시료, 4종의 젖산균을 혼합 접종한 시효들의 발효온도에 따른 pH 및 산도 변화 경향은 서로 매우 유사하며, 특히 1$14^{\circ}C$에서 이러한 경향은 더 뚜렷하게 나타났다. Lac. plantarum이나 Ped. acidilactici 그리고 Lac. brevis를 접종한 시료들은 control group이나 Leu. mesenteroides, 4종의 젖산균을 혼합접종한 시료보다 더 느리게 pH 및 총산도가 변화하였으며 특히 $14^{\circ}C$에서는 control group과 Leu. mesenteroides 그리고 4종의 젖산균을 혼합 접종한 시료에서만이 발효가 진행되었다. 휘발성산과 비휘발성산의 변화 양상도 control group, Leu. mesenteroides를 접종한 시료, 4종의 젖산균을 혼합 접종한 시료에 있어서 서로 매우 유사하였다. 검출한 휘발성 유기산으로는 formfic, acetic acid뿐이었으며, 이들은 Leu. mesenteroides나 Lac. brevis 등에 의해 주로 생성됨을 알 수 있다. Leu. mesenteroides를 접종한 시료에서 다른 발효온도에서보다 $14^{\circ}C$에서 acetic acid의 함량이 높았다. 비휘발성 유기산은 모든 시료에서 lactic acid가 검출되었고, Leu. mesenteroides를 접종한 시료에서 $21^{\circ}C$$14^{\circ}C$에 citric aicd가 검출되었으며 succinic acid는 controlgroup과 Lac. brevis를 접종한 시료에서 검출되었다.

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Characterization of the ${\beta}-Cyclodextrin$ Glucanotransferase Bacillus firmus var. alkalophilus and Its Expression in E. coli

  • Park, Tae-Hyung;Shin, Hyun-Dong;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • 제9권6호
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    • pp.811-819
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    • 1999
  • The ${\beta}-CGTase$ gene of alkalophilic Bacillus firmus var. alkalophilus was cloned into E. coli using $pZErO^{TM}-2$ as a vector. The cloned gene encoded a total of 710 amino acid residues consisting of 674 amino acids of the matured protein and 36 amino acids of the signal peptide, including 20 amino acids from the lacZ gene in the vector. Although the cloned ${\beta}-CGTase$ gene did not contain the promoter and start codons, it was expressed by the lac promoter and lacZ start codon in the $pZErO^{TM}$ vector. A comparison was made with the amino acid sequence and ten other CGTases from Bacillus sp. Also, ten highly conserved regions, which are important amino acid residues in catalysis of CGTase, were identified. The lac promoter used for expression of the ${\beta}-CGTase$ gene was induced constitutively in recombinant E. coli even without IPTG possibly because of a lack of the lacI gene in both host and vector, repressing the lacZ gene in the lac operon. Its expression was catabolically repressed by glucose, however, its repression was reduced by soluble starch, mainly because of the extremely high increase of the cAMP level. ${\beta}-CGTase$ can be overproduced in the recombinant E. coli by maintaining intracellular cAMP levels mostly through the intermittent feeding of glucose during cultivation.

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An Interesting Story of Four Gamma-ray Bright AGNs by the iMOGABA

  • Lee, Sang-Sung
    • 천문학회보
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    • 제43권1호
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    • pp.37.1-37.1
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    • 2018
  • A Korean VLBI Network key science program, the Interferometric Monitoring of Gamma-ray Bright AGNs (iMOGABA) program continues to reveal the nature of the gamma-ray flares in active galactic nuclei (AGNs). Here in this presentation, we would like to introduce an interesting story about four gamma-ray bright AGNs - BL Lac, 1633+382, 3C 84, and M87 - based on the recent results of the iMGOABA. The results will include a) a sad story of an 'orphan' gamma-ray flare from BL Lac, b) a position offset of 40 pc for a gamma-ray flaring site from the radio regions in 1633+382, c) a position alignment of a gamma-ray flaring site with a central engine region in 3C 84, and d) a flat millimeter spectrum of a core in M87 revealed by the iMOGABA.

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Ginsenoside Rb1 exerts neuroprotective effects through regulation of Lactobacillus helveticus abundance and GABAA receptor expression

  • Chen, Huimin;Shen, Jiajia;Li, Haofeng;Zheng, Xiao;Kang, Dian;Xu, Yangfan;Chen, Chong;Guo, Huimin;Xie, Lin;Wang, Guangji;Liang, Yan
    • Journal of Ginseng Research
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    • 제44권1호
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    • pp.86-95
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    • 2020
  • Background: Ginsenoside Rb1 (Rb1), one of the most abundant protopanaxadiol-type ginsenosides, exerts excellent neuroprotective effects even though it has low intracephalic exposure. Purpose: The present study aimed to elucidate the apparent contradiction between the pharmacokinetics and pharmacodynamics of Rb1 by studying the mechanisms underlying neuroprotective effects of Rb1 based on regulation of microflora. Methods: A pseudo germ-free (PGF) rat model was established, and neuroprotective effects of Rb1 were compared between conventional and PGF rats. The relative abundances of common probiotics were quantified to reveal the authentic probiotics that dominate in the neuroprotection of Rb1. The expressions of the gamma-aminobutyric acid (GABA) receptors, including GABAA receptors (α2, β2, and γ2) and GABAB receptors (1b and 2), in the normal, ischemia/reperfusion (I/R), and I/R+Rb1 rat hippocampus and striatum were assessed to reveal the neuroprotective mechanism of Rb1. Results: The results showed that microbiota plays a key role in neuroprotection of Rb1. The relative abundance of Lactobacillus helveticus (Lac.H) increased 15.26 fold after pretreatment with Rb1. I/R surgery induced effects on infarct size, neurological deficit score, and proinflammatory cytokines (IL-1β, IL-6, and TNF-α) were prevented by colonizing the rat gastrointestinal tract with Lac.H (1 × 109 CFU) by gavage 15 d before I/R surgery. Both Rb1 and Lac.H upregulated expression of GABA receptors in I/R rats. Coadministration of a GABAA receptor antagonist significantly attenuated neuroprotective effects of Rb1 and Lac.H. Conclusion: In sum, Rb1 exerts neuroprotective effects by regulating Lac.H and GABA receptors rather than through direct distribution to the target sites.

황소개구리(Rana catesbeiana) 조직의 젖산탈수소효소의 역학적 특성 (Kinetic Properties of Lactate Dehydrogenase in Tissues from Rana catesbeiana)

  • 염정주;하은성
    • 생명과학회지
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    • 제24권2호
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    • pp.118-127
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    • 2014
  • 황소개구리(Rana catesbeiana) I (2월)과 II (8월) 조직의 젖산탈수소효소(EC. 1.1.1.27, lactate dehydrogenase, LDH) 동위효소의 발현 및 역학적 특성을 확인하였다. I의 골격근에서 LDH 활성, $A_4$ 동위효소 및 LDH/CS (EC 4.1.3.7, citrate synthase)가 높게 측정되었고, II의 여러 조직에서 LDH $B_4$ 동위효소의 활성이 증가되었으며, 특히 심장과 뇌조직에서 LDH 활성이 크게 확인되었다. 면역침강반응 후 native-PAGE에 의해 눈조직에서 LDH eye-specific C 동위효소가 확인되었고, LDH C가 LDH B에 유사한 것으로 확인되었다. LDH $A_4$ 동위효소를 oxamate-linked affinity chromatography로 정제하였고, 하부단위체 A의 분자량은 32.0 kDa이었다. II의 LDH는 $Km^{PYU}$이 높았고, 심장과 뇌조직의 $Vmax^{PYU}$이 높았으며, 조직들의 $Vmax^{LAC}$도 높고, 젖산에 대한 내성이 큰 것으로 확인되었다. 그리고 정제한 $B_4$ 동위효소와 눈조직 LDH가 젖산에 대한 내성이 가장 크게 확인되었다. $Km^{LAC}$$Km^{PYU}$보다 컸다. 피루브산 10 mM에 의해 I, II 조직 LDH의 하부단위체 B의 비가 증가함에 따라 LDH 활성의 억제정도가 높았다. 실험 결과, 황소개구리에서 LDH eye-specific C가 확인된 것이 특징적이었고, 황소개구리 I의 골격근 LDH의 활성이 높아 혐기적 대사가 우세하였으며, 황소개구리 II의 경우 LDH B가 증가되고 젖산에 대한 내성이 커져 잘 적응되어진 것으로 사료된다.

The Study of Mutation Spectrum in Iac / Gene of Transgenic Big Blue$\textregistered$ Cell Line Following Short-Term Exposure to 4-Nitroquinoline N-oxide

  • Youn, Ji-Youn;Kim, Kyung-Ran;Cho, Kyung-Hea;Ryu, Jae-Chun
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 1996년도 제19회정기학술대회(The 19th Symposium of the Korean Society of Environmental Toxicology)
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    • pp.64-64
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    • 1996
  • Transgenic animal and cell line models which are recently developed in toxicology field combined with molecular biological technique, are powerful tools for studying of mutation in vivo and in vitro, respectively. The Big Blue mutagenesis assay system is one of the most widely used transgenic systems. Especially, for the study of direct acting mutagens, Big Blue cell line is very useful and powerful to evaluate mutagenicity because the mutation frequency and mutationspectrlun showed no distinct differences between cell line and animal. The Big Blue cell lines carry stably integrated copies of lambda shuttle vector containing lac I gene as a mutational target. These lambda shuttle vectors are useful for various mutagenesis related studies in eukaryotic system due to their ability to be exposed mutagen and then transfer a suitable target DNA sequence to it convenient organism for analysis. We tried to assess the mutagenic effect of 4-NQO with Big Blue cell line. After the treatment of 4-NQO, genomic DNA was isolated and lambda shuttle vector was packaged by in Vitro packaging and then these were plated on bacterial host in the presence of X-gal to screen mutation in the lac I. We determined MF as a ratio of blue plaques versus colorless plaques and now undergoing the mutation spectrum of 4-NQO in lac J gene sequence.

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Impacts of sintering temperature on the mechanical properties and gamma ray protection capacity of clay bricks

  • Aljawhara H. Almuqrin;K.A. Mahmoud;M.I. Sayyed;H. Al-Ghamdi
    • Nuclear Engineering and Technology
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    • 제56권7호
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    • pp.2489-2497
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    • 2024
  • The current study aims to develop new clay bricks doped with metallic waste for radiation shielding applications. The aforementioned new bricks were fabricated with various metallic waste concentrations under a pressure rate reaching ≈114 MPa and firing temperature of 1100 ℃. The impacts of the metallic waste and the firing temperature on the developed brick samples' physical, radiation shielding, and structural properties were studied. In order to identify the fabricated bricks' mineral content, the X-ray diffraction pattern was used. Additionally, the fabricated bricks' porosity and density were experimentally determined, where the porosity was reduced by 28.03%, while their densities increased by ≈ 10.5% by raising the concentration of metallic waste. The linear attenuation coefficient (LAC) for the developed brick was investigated experimentally using a NaI (Tl) scintillation detector over the 0.033-1.408 MeV energy interval. The measured LAC values were enhanced by increasing the concentrations of metallic waste within the fabricated bricks over the examined energy interval. The fabricated brick's LAC enhancement improves the gamma-ray shielding characteristics. Therefore, the fabricated bricks are a cheap and suitable choice for radiation protection applications.