• Title/Summary/Keyword: -galactosidase

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Retroviral Gene Expression in Spermatogonial Stem Cells during Long-term Culture

  • Jeong, Dong Kee;Griswold, Michael D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.7
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    • pp.1015-1022
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    • 2007
  • The spermatogonial stem cell (SSCs) is unique in that it is the only cell in the adult male that can contribute genes to a subsequent generation. Permanent modification of the germ cell line may be realized if stem cells could be cultured, transfected with unique genes, and then transplanted into recipient testes. We developed a culture system that supported long-term viability of SSCs. We used a retrovirus vector (pMSCV including ${\beta}$-galactosidase) to stably transfect spermatogonia following long-term culture using the system developed. Expression of the reporter gene ${\beta}$-galactosidase controlled by the retroviral vector was stable in long-term cultured SSCs. We confirmed the retroviral-mediated ${\beta}$-galactsidase gene could be expressed in germ cells in recipient mice following SSCs transplantation.

Antimicrobial Activities of Korean Medicinal Herb Extracts for Preserving Greenhouse Fresh Produce (시설채소산물의 선도유지를 위한 한국산 약용식물추출물의 항균특성)

  • 정순경;이숙지
    • Food Science and Preservation
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    • v.5 no.1
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    • pp.13-21
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    • 1998
  • Korean medicinal herb extracts(KMHE) were applied to the preservation of greenhouse produce in order to prove their effectiveness. KMHE showed remarkable antimicrobial effects against Bacillus cereus, Peudomonas syringae, and Corynebacterium xerosis causing the postharvest decay of greenhouse produce. Among KMHE the extracts of Rheum palmatum L. and Coptis chinensis Franch most obviously inhibited the growth of microorganims causing the Postharvest decay of greenhouse produce, which destroyed to undetectable levels when treated with more than 500ppm of KMHE. The activities of KMHE were stable in the wide spectrum of pH and temperature. Direct visualization of microbial cells by using both transmission electron microscope and scanning electron microscope showed microbial cell membrane the function of which was destroyed by treating with the dilute solutions of KMHE. This change of cellular membrane permeability could be identified in the experiment that O-nitrophenyl-$\beta$-D-galactopyranoside(ONPG), the artificial substrate of $\beta$-galactosidase, was hydrolyzed in the presence of KMHE, indicating that the membrane was perturbed by KMHE.

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EARLY SCREENING OF EXPRESSION OF SV40 DRIVEN LACZ INTRODUCED INTO BOVINE EMBRYOS

  • Nakamura, A.;Okumura, J.;Muramatsu, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.5
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    • pp.449-454
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    • 1995
  • The present study was conducted to assess gene expression of bacterial lacZ driven by the SV40 promoter at early developmental stages of bovine embryos. The lacZ gene was linearized with BamHI digestion and introduced into the pronucleus by microinjection at 20 hrs after the commencement of in vitro fertilization. Intact bovine blastocysts were not stained with X-Gal, suggesting that there is no endogenous beta-galactosidase activity in these blastocysts. In contrast, the bovine blastocyst cells microinjected with the lacZ gene exerted a characteristic greenish-blue color originating from the bacterial beta-galactosidase activity, albeit at a low rate, i.e. 2.1% of the total fertilized oocytes injected. It was concluded, therefore, that the lacZ gene driven by the SV40 promoter could be used for an indirect screening method in which the presence of transgene is evaluated from the product of transgene expression.

Mutational Analysis of the Region between vir Box and -35 Sequence in virE Promoter of pTiA6

  • Woong Seop Sim
    • Journal of Plant Biology
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    • v.38 no.3
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    • pp.259-266
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    • 1995
  • To investigate how the dyad symmetry region (DSR) and the distance between vir box and -35 sequence of the virE promoter plays a role in virE gene expression, two mutants were constructed by base substitution and insertional mutagenesis. The base substitutional mutation, a AAlongrightarrowCG substitution at positions -39 and -40 on the DSR, showed the level of $\beta$-galactosidase activity approximately 91% of the wild type virE promoter activity. Therefore, the native structure of the DSR seems to be not essential for virE expression. The insertional mutation, constructed by inserting 8 bp ClaI linker between -49 and -50, displayed the $\beta$-galactosidase activity at 12% of the native virE promoter activity. However, this striking reduction appears to be not caused by destruction of the native DSR structure, but by shifting the vir box far from putative -35 sequence.

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Isolation of N-Containing Sugars from Silkworm Urine and Their Glycosidase Inhibitory Activities (잠뇨로부터 질소함유 당물질 분리 및 glycosidase에 대한 저해활성)

  • 송주경;정성현
    • Biomolecules & Therapeutics
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    • v.6 no.4
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    • pp.364-370
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    • 1998
  • Glycosidase inhibitors from urine of Bombyx mori were isolated and their inhibitory activities on glycosidases were evaluated. Six compounds were isolated by using several ion exchange columns, and their chemical structures were identified by the physicochemical and spectral data. Compound IV, V and Ⅵ were identified as 1-deoxynojirimycin, fagomine and 1,4-dideoxy-1,4-imino-D-arabinitol, respectively. Among six compounds isolated,1-deoxynojirimycin(IV) was the most potent inhibitor on $\alpha$-glucosidase and $\beta$-galactosidase of rat intestine, and its inhibitory activities for trehalase and almond $\beta$-glucosidase were relatively weak. Compound V and Ⅵl retained a little inhibitory potency toward $\alpha$-glucosidase and $\beta$-galactosidase. Compound II and III, however, have been found to have no effect on all glycosidases tested in this study.

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Type 2 GM1 Gangliosidosis Presented with Developmental Regression: A Case Report (발달의 퇴행을 보여 진단된 제2형 GM1 gangliosidosis 1례)

  • Ko, Jung Min;Cho, Tae Joon;Chae, Jong-Hee
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.14 no.2
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    • pp.182-185
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    • 2014
  • GM1 gangliosidosis is a rare autosomal recessively inherited metabolic disease due to deficiency of ${\beta}$-galactosidase caused by mutations in the GLB1 gene. There have been three clinical subgroups in GM1 gangliosidosis, however it is difficult to differentiate because there is considerable overlap between classical phenotypes and clinical and imaging findings among the subgroups. Here, we report a Korean girl with type 2 GM1 gangliosidosis, who showed dysostosis multiplex and progressive neurological deterioration. Developmental regression was first noted at the age of 9 months, and she was diagnosed as GM1 gangliosidosis by ${\beta}$-galactosidase enzyme analysis and GLB1 mutation analysis at the age of 16 months.

Regulation of fpr Gene Encoding NADPH : Ferredoxin Oxidoreductase by the soxRS Locus in Escherichia coli

  • Koh, Young-Sang;Choih, Jenny;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.137-143
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    • 1996
  • We isolated a promoter inducible by paraquat, a superoxide-generating agent, from Escherichia coli using a promoter-probing plasmid pRS415. From sequence analysis we found out the promoter is for fpr ENCODING nadph : ferredoxin oxidoreductase. We constructed on operon fusion of lacZ gene with fpr promoter to monitor the expression of the gene in the single-copy state. LacZ expression generators, menadione and plumbagin, also induced the expression of .betha.-galactosidase in the fusion strain. On the other hand, no significant induction was observed by treatment with hydrogen peroxide, ethanol, and heat shock. Induction of .betha.-galactosidase was significantly reduced by introducing a .DELTA. sox 8 :: cat of soxS3 :: Tn10 mutation into the fusion strain, indicating that fpr gene is a member of the soxRS regulon. The transcriptional start site was determined by primer extension analysis. Possible roles of fpr induction in superoxide stress were discussed.

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Changes in the Cell Wall Components, Enzyme Activities and Cell Structure of Peach during Maturation (복숭아의 품종과 숙도에 따른 세포벽 성분, 효소활성 및 조직의 변화)

  • 김미현;신승렬
    • Food Science and Preservation
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    • v.1 no.2
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    • pp.107-116
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    • 1994
  • This paper was carried out to Investigate changes in the activities of cell-degrading enzymes, cell wall components and cell structure of peach during maturation and storage for valuation of quality. The firmness of peach was decreased during maturation and storage, and was remarkably decreased in Daegubo than Yumyung. Polygalacturonase and $\beta$-galactosidase activities of peach were increased during maturation and storage, and were remarably increased in soft peach and in mature and soft peach, respectively. Contents of alcohol-insoluble substance, cell wall, and total and insoluble pectin of peach were decreased during maturation and storage, but cellulose and soluble pectin were increased. Intracellular space was enlarged during maturation and middle lamella was gradually degraded during maturation.

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Studies on the lactose constitutive mutants of Streptococcus lactis $KB_{21}$ (Streptococcus lactis $KB_{21}$의 lactose constitutive mutant에 관한 연구)

  • Park, Yun-Hee;Mckay, Larry L.
    • Applied Biological Chemistry
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    • v.23 no.4
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    • pp.218-221
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    • 1980
  • From S. lactis $KB_{21}$, stabilized strain by intergration of the lactose plasmid into the host chromosome, several lactose constitutive mutants were obtained by UV irradiation and spontaneous mutation. The rapid growth of the mutants in the medium containing lactobionate confirmed their constitutive nature. The mutants synthesized $phospho-{\beta}-galactosidase$ with an activity of 1.7 to 3.4 times that of the parent.

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Inhibition Mechanism of $\alpha$-D-Glucosidase Inhibitor from Streptomyces sp (Streptomyces속 균주가 생성하는 $\alpha$-D-Glucosidase 저해물질의 작용상)

  • 도재호;주현규
    • Microbiology and Biotechnology Letters
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    • v.18 no.1
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    • pp.39-43
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    • 1990
  • The inhibitor had the inhibitory activities against hydrolysis of PNPG, sucrose and ONPG by $\alpha$-Dglucosidase, $\alpha$ - and $\beta$ -galactosidase, but it did not inhibit amylases and other carbohydrases. Kinetic studies exhibited that the inhibitory substance non-competitively inhibited the enzyme reaction with a Ki value of 118 $\mu$g/m$\ell$, and enzyme-inhibitor complex was formed slowly.

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