• Archives of Pharmacal Research
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• 제26권5호
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• pp.367-374
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• 2003

In the course of screening neuraminidase inhibitors from herbal medicines, Reynoutria elliptica exhibited high inhibitory activity. Four active compounds were isolated from the ethyl acetate soluble fraction by consecutive purification using sillica gel, Sephadex LH-20 chromatography, and recrystallization. The chemical structures of these compounds were identified as 1,3,8-trihydroxy-6-methylanthraquinone (emodin) 1,8-dihydroxy-3-methoxy-6-methylanthraquinone (emodin 3-methyl ether; physcion), 1,3,8-trihydroxy-6-hydoxymethylanthraquinone ($\omega$-hydroxyemodin), and 3,5,4 -trihydroxystilbene (trans-resvertrol) by spectral data including MS, $^1 H-, and ^{13}C-NMR. The IC_{50}$ values of emodin, emodin 3-methyl ether, $\omega$-hydroxyemodin, and trans-resvertrol were 2.81, 74.07, 10.49, and 8.77 $\mu$M, respectively. They did not inhibit other glycosidase such as glucosidase, mannosidase, and galactosidase, indicating that they were relatively specific inhibitors of neuraminidase.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1993년도 제2회 신약개발 연구발표회 초록집
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• pp.174-174
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• 1993

목적: Cinnamic acid의 유도체들중에서 항돌연변이 효과가 보고된 바 있어, 본 연구에서는 E. coil PQ37균주를 사용한 SOS chromotest를 이용하여 약 170개 cinnamic acid의 유도체들의 항돌연변이 효과를 확인하고자 하였고, 1차로 확인된 수종의 유도체들에 대한 결과를 보고하고자 한다. 방법: SOS chromotest는 sul A:: lacz fusion strain 인 E. coli PQ37 에서의 chemical로 인한 DNA damage에 대한 SOS response를 $\beta$-galactosidase enzyme activity level로서 측정하는 실험이며, m-RNA 또는 protein synthesis에 대한 test chemicals의 cytotoxic저해효과를 알아보기 위하여 alkaline phosphatase를 병행측정하여 보완하였다. 결과 및 고찰: (－)S－9 경우는 4-nitroquinoline 1-oxide(4-NQO)를 유도물질로 하였으며 대략 Induction factor가 9.7 정도였고, (＋)S－9의 경우는 aflatoxinB$_1$을 유도물질로 하였고 이때의 Induction factor는 13.8 정도였다. 이결과 1차로 test된 cinnamic acid유도체 6개중에서 RK001, RK002, RK003, RK004, RK005는 거의 항돌연변이 효과를 나타내지 않았으나, RK006은 항돌연변이 효과를 보여주어 이들의 Structure-Activity Relationship (SAR) 및 Dose-response와 RK006의 항돌연변이 효과의 mechanism에 관해 M13 mp2 viral DNA의 유전자 염기서열을 이용하여 항돌연변이 기전연구를 수행중이다.

• 한국가축번식학회지
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• 제17권3호
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• pp.263-267
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• 1993

Transgenic animals have become an important tool in the basic and applied sectors of genetic and biomedical sciences. In particular transgenes provide clear-cut markers in the spatial and temporal analysis of developing embryos for the understanding of developmental mechanisms. For the long-term use of plasmid vector in a particular purpose it would be necessary to develop one's own vector system which can be properly expressed in eukaryotic system. Plasmids were constructed from ori region of pUC19 and early region of SV40 through various steps. LacZ gene coding for $\beta$-galactosides was fused to early gene of SV40 in translational in-frame. Poly(A) tailing site of SV40 was inserted at the 3' lacZ so that initiation, elongation and terminatin be controlled by SV40 transcription (pSS4). Biological function of the constructed pSS4 was demonstrated via microinjection of the plasmid into fertilized loach eggs and subsequent detection of $\beta$-galactosidase in developing embryos. The result indicate that the newly constructed pSS4 is functional in a eukaryotic system in vivo. Thus pSS4 may be used as an efficient tool for the study of embryogenesis and a basic carrier for various genes for animal transgenesis.

• Food Science and Biotechnology
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• 제18권1호
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• pp.172-178
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• 2009

Fermented ginseng (FG) is an ethanol extract of ginseng radix processed with $\beta$-galactosidase. It was hypothesized that FG may exert anti-hyperlipidemic and anti-diabetic activities through modulating AMP-activated protein kinase (AMPK) in HepG2 human hepatoma cells. In this study, we showed that AMPK phosphorylation was stimulated by FG. These effects were abolished by pretreatment with an AMPK inhibitor, compound C. In addition, FG regulated the expression of genes associated with lipogenesis and lipolysis, thus causing suppression of hepatic triglyceride accumulation. In vivo study using db/db mice, FG reduced fasting plasma glucose, HbAlc, and insulin resistance index, when compared to diabetic control. FG also increased the phospho-AMPK and glucose transporter 4 (GLUT4) expressions in liver and skeletal muscle, respectively. In liver, expressions of lipogenic gene were decreased whereas expressions of lipolytic genes were induced, when compared to diabetic control. Taken together, we may suggest that FG ameliorates hyperglycemia and hyperlipidemia through activation of AMPK and could be developed as a health functional food or therapeutic agent for type 2 diabetic patients.

• Journal of Microbiology and Biotechnology
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• 제5권5호
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• pp.245-249
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• 1995

Expression of the adjacent but divergently transcribed glpD and glpE gene is positively regulated by cAMP-CRP. In this study, we constructed several mutants in which a CRP-binding site is placed at different distances upstream of the glpD promoter. The effect of the spacer length on transcription activation by cAMP-CRP was tested in vivo by $\beta$-galactosidase. The cAMP-CRP complex activated transcription from glpD when bound at a number of positions, all of which lay on the same face of the DNA helix, although the degree of activation varied with the length of the spacer. By contrast, the insertion of spacer length with non-integral turns of the DNA helix extremely inhibited the activation of transcription. The observed transcription activation by cAMP of the glpD promoter was influenced by the distance between the CRP binding site and the transcription start point.

• 약학회지
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• 제55권1호
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• pp.56-63
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• 2011

Fabry disease (FD) is an X-linked inborn error of glycoshpingolipid metabolism resulting from mutation in the enzyme ${\alpha}$-galactosidase A gene. The disease is an X-linked lipid storage disorder and the lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly globotriaosylceramide (Gb-3). Measurement of Gb-3 in plasma has clinical importance for monitoring after enzyme replacement therapy for confirmed FD patients. Using electrospray ionization MS/MS we had developed, a simple, rapid, and highly sensitive analytical method for Gb-3 in plasma was used for the purpose of screening FD among high risk groups in Korean population. To date, no comprehensive results for FD screening have been performed and reported in Korea. We screened 1,100 outpatients from 13 hospitals (including clinics) to assess the incidence of FD among patients in high risk groups. For patients with borderline level amount of Gb-3, we repeated Gb-3 or performing complementary or confirmative assay with ${\alpha}$-Gal A activity and DNA mutaion analysis for confirmation diagnosis. Of 1,100 we diagnosed 3 FD with 2 classical type and 1 carrier (0.27%).

• Animal cells and systems
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• 제1권4호
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• pp.637-640
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• 1997

Raf-1, a cytoplasmic serine/threonine protein kinase, serves as a central intermediate in many signaling pathways in cell proliferation, differentiation, and development. In this study, we investigated that B-raf, Drosophila homolog of the human c-raf-1, is involved in ultraviolet (UV) responsive events by using hypomorphic mutant $D-raf^{c110}$ and Draf-lacZ transgenic fly. At first, effect of UV damage on the survival of wild-type and $D-raf^{C110}$ strains was examined. In terms of $1/LD_{50}$ value, the relative ratio of UV sensitivities of wild-type versus $D-raf^{C110}$ strain was 1 : 2.2. By using quantitative $\beta$-galactosidase activity analysis, transcriptional activity of the D-raf gene promoter was also examined in UV-irradiated Draf-lacZ transgenic larvae. UV irradiation increased the expression of lacZ reporter gene in Draf-lacZ transgenic fly. However, in $D-raf^{C110}$ strain the transcriptional activity of D-raf gene promoter by UV irradiation was extensively reduced. Results obtained in this study suggest that D-raf plays a role in UV response, leading to better survival of Drosophila to UV damage.

• 한국동물학회지
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• 제38권2호
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• pp.196-203
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• 1995

The c-myc proto-oncogene, one of the immediately earlY genes, is expressed in various mammalian cell types and heavily involved in the regulation of cell proliferation and differentiation. To determine endogeneous expression pattern of c-myc gene in preimpBantation mouse embwos, we employed a reverse transcription coupled to polvrnerase chain reaction (RT-PCR). Transcript of c-myc was detected at fertilized embryos as a maternal transcript. At the early two-cell stave, transcript of c-myc gene was hardly detected, bu, appeared at late two-cell embryos as a zygotic transcript. The level of c-myc expresion was increased at later stases and peaked at blastocvst stage. To examine the functional role of promoter region for c-myc gene transcription, we fused the 5'upstream region (1.8 kb) including econ 1 of c-myc genomic DNA with E. coli lacE gene fnamed as pcMYC-laczl. pcMYC-lacZ was microiniected into the pronscleus of mouse one-cell embryovs, and p·salactosidase activity was determined tv histochemical staining with X-gal at different stases. f-galactosidase activity was detected only at blastocyst, but not at the earlier stage embryos. This result indicates that c-myc gene is transcriptionallv active during mouse preimplantation development.

• 미생물학회지
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• 제31권1호
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• pp.27-36
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• 1993

Rhizobium fredii USDA191 은 대기 중의 질소를 환원하여 식물체의 생육에 필요한 질소원을 공급해주는 세균으로 다량의 체외 다당류를 합성한다. 전위요소 Tn5의 삽입에 의한 돌연변이 유도로 다당류결핍 변이주 R. fredii YKL293 가 분리되었으며 이 변이주로부터 Tn5 에 인접한 DNA 단편이 pUC19 에 클로닝되었고(plyk5293),이 DNA 단편을 탐침으로 하여 .lambda.NM1149 에 구성되 USDA191 genomic library 로부터 야생형체외다당류 합성관련 유전자(exo) 를 함유한 클론 .lambda. NM1149 22E 를 plaque 혼성화에 의하여 분리하였다. 클론 NM1149.22E 에 들어있는 exo 유전자를 pBR322 에 옮겨서 pJW33을 만들고, 재조합체 pJW33 을 Escherichia coli POII734 에 도입시켜 lacZ 구조유전자를 함유한 MudI 1734 가 exo 유전자의 프러모토와 융합되어 lacZ 구조유전자의 전사가 이루어지도록 하였다. 위와 같이 만들어진 재조합체 플라스미드 pUM21을 함유한 E. coli JM83 은 .betha.-galactosidase 를 합성하였으며, 야생형 tacZ 유전자를 갖고 있는 E. coli LE392 에 비해서 14-25배 정도 낮은 역가를 보였다.

• Journal of Microbiology and Biotechnology
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• 제9권4호
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• pp.393-397
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• 1999

The gene (galE) encoding UDP-galactose-4-epimerase, operative in the galactose metabolic pathway, was cloned together with the $\beta$-galactosidase gene (lacZ) from Lactococcus lactis ssp. lactis ATCC7962 (L. lactis 7962). galE was found to have a length of 981 bps and encoded a protein with a molecular mass of 36,209 Da. The deduced amino acid sequence showed a homology with GalE proteins from several other microorganisms. A Northern analysis demonstrated that galE was constitutively expressed by its own promoter. When galactose or lactose was added into medium, the galE transcription was induced by several upstream promoters. The structure of the gal/lac operon of L. lactis 7962 was partially characterized and the gene order around galE was galT-lacA-lacZ-galE-orfX.