• Title/Summary/Keyword: lac I

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

  • Ryu, Jae-Chun;Youn, Ji-Youn;Cho, Kyung-Hae;Chang, Il-Moo
    • Environmental Mutagens and Carcinogens
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    • v.18 no.1
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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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Extracellular Production of Alpha-Interferon by Recombinant Escherichia coli: Part II. The Growth Behavior of the Recombinant Cells (유전자 재조합 대장균을 사용한 Alpha-interferon의 생산과 분비: 제2부. 재조합 균주의 생장특성)

  • 노갑수;최차용
    • KSBB Journal
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    • v.5 no.3
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    • pp.195-200
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    • 1990
  • The growth behavior of recombinant Escherichia coli cells having plasmid pIF-III-B, which carries human alpha-interferon gene under the control of lpp promoter, lac promoter and lac operator, was studied by using of various E. coli host strains. Expression of the alpha-IFN gene is controllable by using inducer IPTG because the plasmid also contains lacI gene which produces lac regressors. The repressors block the transcription of alpha-IFN gene. There were considerable differences in cell growth according to the host strains used. Cell growth was inhibited not only by plasmid pIF-III-B itself but also by the induction of alph-a-IFN gene expression. Growth inhibition caused by the plasmid itself was more serious than that caused by the induction of alpha-IFN gene expression.

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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
    • Journal of the Korean Magnetic Resonance Society
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    • v.13 no.2
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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.

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

  • Cho, Young;Rhee, Hei-Soo
    • Korean journal of food and cookery science
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    • v.7 no.1
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    • pp.15-25
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    • 1991
  • The effects of lactic acid bacteria on the chemical and microbial changes of fermented kimchi at various temperatures were studied. Kimchi was homogenized and was sterilized by ultra violet, then Lactobacillus plantarum, Leuconostoc mesenteroides, Pediococcus acidilactici, Lactobacillus brevis and the mixture of there bacteria inoculated on sterilized kimchi, respectively. The measurement of pH and total acidity, quantitative analysis of volatile organic acids and nonvolatile organic acids by gas chromatography were investigated while inoculated kimchi were fermented at $30^{\circ}C$, $21^{\circ}C$, $14^{\circ}C$ and $7^{\circ}C$. Sample I (original kimchi homogenate), Sample III (inoculated with Leuconostoc mesenteroides) and Sample Ⅵ (inoculated with mixed lactic acid bacteria) were alike in that changes of pH and total acidity and especially, these phenomena were prominent at $14^{\circ}C$. Formic, acetic and heptenoic acid as volatile organic acid were detected by GC, and these acids formed mainly by Leuconostoc mesenteroides and lactobacillus brevis. Sample III was more higher content than other samples at $14^{\circ}C$. As nonvolatile organic acid, lactic acid in all samples, citric acid in sample III at $21^{\circ}C$and $14^{\circ}C$, succinic acid in sample I at $30^{\circ}C$, $21^{\circ}C$, $14^{\circ}C$ and sample V at $30^{\circ}C$ were detected by GC.

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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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    • v.9 no.6
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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
    • The Bulletin of The Korean Astronomical Society
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    • v.43 no.1
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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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    • v.44 no.1
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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.

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

  • Yum, Jung Joo;Ha, Eun Sung
    • Journal of Life Science
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    • v.24 no.2
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    • pp.118-127
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    • 2014
  • The kinetic properties and isozyme expression of lactate dehydrogenase (EC 1.1.1.27; LDH) in tissues from Rana catesbeiana I and II collected from February (I) and August (II) were studied. LDH activities, A4 isozyme, and LDH/citrate synthase (EC 4.1.3.7; CS) were high in skeletal muscle from R. catesbeiana I, and LDH $B_4$ isozyme increased in several tissues of R. catesbeiana II. In particular, LDH activities were high in heart and brain tissues from R. catesbeiana II. LDH eye-specific C isozyme, detected by native polyacrylamide gel electrophoresis after immunoprecipitation, was expressed in eye tissue and was more similar to the $B_4$ than $A_4$ isozyme. LDH $A_4$ isozyme was purified by oxamate-linked affinity chromatography, and the molecular weight of subunit A was 32.0 kDa. In R. catesbeiana II, levels of $Km^{PYU}$, $Vmax^{LAC}$, and tolerance to lactate of LDH were high in all tissues, and $Vmax^{PYU}$ of LDH in heart and brain tissue was highly detected. Purified $A_4$ isozyme and LDH in eye tissue were highly tolerate compared to others. The $Km^{LAC}$ value was highly measured compared to $Km^{PYU}$. The degree of inhibition by 10 mM of pyruvate on LDH activities in tissues from R. catesbeiana I and II was more pronounced as the ratio of subunit B increased. As a result, characteristic expression of LDH eye-specific C was found in R. catesbeiana. Anaerobic metabolism seemed to predominate as the LDH of skeletal muscle from I showed higher activity. It also appeared that R. catesbeiana II adapted well to incremental increases in LDH B, becoming tolerant to the lactate of LDH in tissues.

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
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 1996.12a
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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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Molecular Cloning of Antagonistic Genes in Pseudomonas maItophiliQ B-14 (토양병해 길항성 Pseudomonas maltophilia B-14의 길항유전자탐색)

  • 구본성;서영우;윤상홍;박경수;은무영;김용환;오상우;류진창;은무영
    • Microbiology and Biotechnology Letters
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    • v.20 no.6
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    • pp.619-624
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    • 1992
  • Tn5 lac 삽입으로 채소입고병원균에 길항력이 약화된 T-67 및 고추역병균과 참깨역병균에 길항력이 약화된 T-81의 Tn5 lac 유전자 일부와 오른쪽 말단에 있는 길항관련 유전자의 flanking sequence가 cloning된 pAG67 및 pAG81 clone을 선발하였고, pAG67 및 pAG81 clone된 길항관련 유전자의 flanking sequence를 야생 길항균 Pseudomonas maltophilia B-14의 DNA를 probe로 사용하여 Southern hybridization으로 확인하였으며, 제한효소 지도를 작성하여 8Kb 및 4Kb 크기의 flanking sequence가 cloning되었음을 확인하였다. pAG6 및 pAG81의 flanking sequence를 EcoRi-BglII와 EcoRI-MpaI으로 분리하여 유전자 은행으로부터 길항관련 유전자가 cloning된 cosmid clone 7개주를 선발하였다.

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