• Title/Summary/Keyword: Apase

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Construction of an Expression Vector System with the GAP Promoter in Saccharomyces cerevisiae (효모, Saccharomyces cervisiae의 GAP 유전자를 이용한 발현 벡터계의 개발)

  • 황요일;서애란;심상국;정동효
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.568-574
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    • 1991
  • The cloned glyceraldehyde-3-phosphate dehydrogenase (GAP) gene of Saccharomyces cereviszae (Holland et al., 1983) has been characterized. Based on the communication, we have also cloned 2.1 kb CAP DNA fragment and modified this fragment as a portable promoter. Two yeast expression vectors, one is YCp type vector being maintained at low copy number (1 or 2) and the other is YEp type vector at high copy number, have been constructed with the GAP promoter and the PH05' gene as a reporter. Our plasrnids were introduc,ed into S. cerevisiae HY-1, which has been improved. The $Trp^+$ transformants expressed APase activity efficiently and showed high level of PH05' transcripts.

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Phosphate and Carbon Source Regulation of Alkaline Phosphatase and Phospholipase in Vibrio vulnificus

  • Oh, Wan-Seok;Im, Young-Sun;Yeon, Kyu-Yong;Yoon, Young-Jun;Kim, Jung-Wan
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.311-317
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    • 2007
  • In this study, the effects of phosphate concentration and carbon source on the patterns of alkaline phosphatase (APase) and phospholipase (PLase) expression in Vibrio vulnificus ATCC 29307 were assessed under various conditions. The activities of these enzymes were repressed by excess phosphate (4 mM) in the culture medium, but this repression was reversed upon the onset of phosphate starvation in low phosphate defined medium (LPDM) containing 0.2 mM of phosphate at approximately the end of the exponential growth phase. The expressions of the two enzymes were also influenced by different carbon sources, including glucose, fructose, maltose, glycerol, and sodium acetate at different levels. The APase activity was derepressed most profoundly in LPDM containing fructose as a sole carbon source. However, the repression/derepression of the enzyme by phosphate was not observed in media containing glycerol or sodium acetate. In LPDM-glycerol or sodium acetate, the growth rate was quite low. The highest levels of PLase activity were detected in LPDM-sodium acetate, followed by LPDM-fructose. PLase was not fully repressed by high phosphate concentrations when sodium acetate was utilized as the sole carbon source. These results showed that multiple regulatory systems, including the phosphate regulon, may perform a function in the expression of both or either APase and PLC, in the broader context of the survival of V. vulnificus.

Agrobacterium-Mediated Transformation on a Plant with Saccharomyces cerevisiae Acid Phosphatse Gene(PHO5) (Agrobacterium을 이용한 Saccharomyces cerevisiae Acid Phosphatse 유전자 (PHO5) 의 식물체로의 도입)

  • Ki yong Kim;Dae yuong Son;Yong Gu Park;Won Il Jung;Jin Ki Jo
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.13 no.3
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    • pp.177-183
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    • 1993
  • This study was conducted to obtain the transformed tobacco plants with S. cerevisiae Acid phosphatase gene(PH05) using Agrobacterium tumefaciens and th confirm plant transformation and gene expression. the results obtained were summarized as follows: APase activity of Saccharomyces cereviase NA 87-11A was remarkably showed up as deep red color when assayed by Tohe and Oshima(1974). PH05 fragment, Apase gene, was obtained from pVC727G and the graphically estimated size was about 1.5kb by agarose gel electrophoresis. The sequencing results of 5'end and 3'end of PH05 using dideoxy chain termination method were coinsided with the full length nucleotide already. pBKJ I vector was constructed by isolation of PH05 fragment from pVC727-1 and pBKSI-1 digesred with Sma I and Xba I. Isolated plasmid from transformed A. tumefaciens with constructed pBKJ I when it was electrophoresed with agarose gel. The dosc of tobacco leaf was cocultivated 재소 transformed Agronacterium tumefaciens. Transformed shoots were selected on kanamtcin-containing MS-n/B medium and they were regenerated. The transgenic tobacco plants were elucidated by isolation of genomic DNA and genomic southern hybridization using ${\alpha}-^{32}P$ labelled PH05 fragments. The PH05 in transformed tobacco plants was expressed in leaf, stem and root, and its APase activity was estimated as deep red color by Tohe method.

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The Production of HBsAg in the Recombinant Yeast Cells (재조합 효모 세포내에서의 간염백신 생산)

  • Park, Cha-Yong;Lee, Hei-Chan
    • Microbiology and Biotechnology Letters
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    • v.14 no.6
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    • pp.455-460
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    • 1986
  • Dane particle was prepared from the plasma of chronic HBsAg carrier with high levels of HBsAg activity. DNA extracted front Dane particle core after a DNA polymerase reaction with $\alpha$-($^{32}$P) dNTP, was identified as HBV DNA by liquid scintillation counter and agarose gel electrophoresis-G.M. counting. To produce Hepatitis B surface antigen for use as a vaccine against Hepatitis B virus infection, yeast strains harboring recombinant plasmid with Apase promoter was used. Recombinant plasmid was construced from pHBV 130 and pAN 82, transformed into E coli, and then transferred into yeast strains. HBsAg was produced by derepression in Burkholder minimal medium with controlled inorganic phosphate concentration. The kinetics of HBsAg production was also investigated. Total HBsAg activity increased rapidly between 3 and 6 hours after transfer to phosphate-free medium and reached a maximum at around 9th hour. The transfer into phosphate-free medium after 6 hours in high phosphate cell growth medium gave maximum activity.

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Identification and molecular characterization of a low acid phosphatase 3 (lap3) mutant based on the screening of an Arabidopsis activation-tagged population

  • Jin, Yong-Mei;Won, So-Youn;Jeon, Hye-Sung;Park, Sang-Ryoung;Kim, Min-Kyun
    • Plant Biotechnology Reports
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    • v.5 no.1
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    • pp.45-51
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    • 2011
  • A low acid phosphatase 3 (lap3) mutant was identified and characterized from an Arabidopsis activation-tagged (Weigel) population. The roots of the lap3 plants showed lower acid phosphatase (APase) activity compared to wild-type ones under low-Pi conditions ($10{\mu}M\;Pi$). Plasmid rescue experiments revealed that the activation-tagging vector was inserted into the intergenic region between At4g31540 and At4g31550 in the Arabidopsis genome. The genotypic segregation of the lap3 mutation was tightly linked with the phenotypic segregation of root APase activity in the prgeny of lap3. The transcript level of the At4g31520 (SDA1: SEVERE DEPOLYMERIZATION OF ACTIN 1), located 7.4 kb from the CaMV 35S enhancers in the lap3 mutant, was significantly reduced compared to that in the wild type. It was speculated that cellular actin polymerization may be involved in Pi acquisition in higher plants.

A Comparison in the Value of the Clinical Assays of Dementia with Normal Geriatrics (정상 노인과 치매 환자간 임상화학적 검사 수치의 비교)

  • 손혜연;조경혜
    • Biomedical Science Letters
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    • v.3 no.2
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    • pp.161-168
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    • 1997
  • The measurement of many biochemical changes that could be appeared as aging will be resulted in the very useful and basal marker to maintain healthy life and to predict the health condition and the retardation of the aging. We classified the geriatrics into the normal and same aged dementia on the based of clinical diagnostic as well as MMSE evaluation. And then we were collected blood samples from both groups and examined 13 biochemical assays to screen any valuable biochemical index for dementia. Among 13 assays, several assay data showed the statistically significant differences between two groups. The values of serum albumin, APase, and GOT were significantly decreased in demented group compared to control. On the other hand, the values of glucose and LDH were increased in dementia compared to control. Additionally, the value of Pi was increased but the values of calcium and GPT were decreased, without significant difference (p>0.05). The values of other assays such as bilirubin, BUN, cholesterol, creatinine and total protein did not show any statistically significant differences. Conclusively it may be suggested that dementia might be resulted in the variable biochemical changes which could be affected by many environmental factors as well as dementia itself.

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An investigation of the effect of epigallocathechin-3-gallate on the renal dipeptidase release

  • Kim, Yu-Jin;Park, Eun-Mi;Yoon, Hyun-Joong;Park, Haeng-Soon
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.317.1-317.1
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    • 2002
  • The action of epigallocatechin-3-gi:lllale (EGCG). polyphenol compound from green lea, on the release pattern of glycosylphosphatidylinositol (GPI)-anchored renal dipeptidase (RDPase) from renal proximal tubules (PTs) was examined. EGCG had a stronger inhibitory effect on the release of RDPase than alkaline phosphatase (APase), another GPI-anchored ectoenzyme used as a reference protein. The effect of EGCG on cell viability as assessed by MTT test was found to be intact, and moreover, was indicative of potent cell activation or proliferation. (omitted)

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Nutritional Status and Related Factors of Residents Aged Over 50 in Longevity AreasI. Anthropometric and Biochemical Nutritional Status (고령인구 비율이 높은 지역 장년, 노년층의 건강.영양상태 및 이에 영향을 미치는 인자에 관한 연구 I. 신체계측, 생화학적 영양상태 (혈청지질, 철분지표, 무기질 등))

  • Choe Jeone-Sook;Paik Hee-Young;Kwon Sung-Ok
    • Journal of Nutrition and Health
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    • v.37 no.9
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    • pp.825-837
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    • 2004
  • Studies on the health problems of the elderly have been increased with increasing life expectancy of Koreans. To assess the health status of people over 50 years old including the elderly (678 free-living people: men (168), women (510)), anthropometric measurements and biochemical nutritional status were analyzed. General characteristics and the prevalences of specific diseases were also examined. The mean anthropometric values for males and females were heights of 162 cm and 149 cm, respectively; and weights of 59 kg and 52 kg, respectively, which were much lower than the Korean standards established in the Korean Recommended Dietary Allowances. The mean BMI, WHR (waist hip ratio), and body fat rate (%), which are obesity indices, were significantly higher in females than in males. As the age went up, height, weight, lean body mass, and mid-arm circumference decreased. Whereas, blood pressure (SBP) for women increased. Mean blood HDL-cholesterol levels of men and women were 46.0 mg/dl and 46.3 mg/dl, respectively; their triglyceride levels were 129.1 mg/d1 and 135.6 mg/dl, respectively, which were not significantly different. Total cholesterol and LDL-cholesterol levels were significantly higher in females than in males. For man there were significantly different in ${\gamma}$-GTP, Na levels by age. As the age went up for women, RBC, hemoglobin, hematocrit, HDL-cholesterol, ${\gamma}$-GTP and $Ca^{2+}$ levels significantly decreased. Levels of total protein and albumin were above the lower normal limit for the majority of the participants. Prevalence of anemia, assessed by hemoglobin using World Health Organization (WHO) definition, was 43.9% for men and 42.1% for women. Also, many of the subjects were out of the normal range in total cholesterol (women), HDL-cholesterol, TAS (total antioxidant status), APase, RBC, hemoglobin, hematocrit levels. Therefore, measures to improve the biochemical nutritional status for the people in rural area are required.d.

Comparative Characteristics of Three Human Embryonic Stem Cell Lines

  • Lee, Jung Bok;Kim, Jin Mee;Kim, Sun Jong;Park, Jong Hyuk;Hong, Seok Ho;Roh, Sung Il;Kim, Moon Kyoo;Yoon, Hyun Soo
    • Molecules and Cells
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    • v.19 no.1
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    • pp.31-38
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    • 2005
  • Human embryonic stem (hES) cells have unique features including unlimited growth capacity, expression of specific markers, normal karyotypes and an ability to differentiate. Many investigators have tried to use hES cells for cell-based therapy, but there is little information about the properties of available hES cell lines. We compared the characteristics of three hES cell lines. The expression of SSEA-1, -3, -4, and APase, was examined by immunocytochemistry, and Oct-4 expression was analyzed by RT-PCR. Differentiation of the hES cells in vitro and in vivo led to the formation of embryoid bodies (EBs) or teratomas. We examined the expression of tissue-specific markers in the differentiated cells by semiquantitative RT-PCR, and the ability of each hES cell line to proliferate was measured by flow cytometry of DNA content and ELISA. The three hES cell lines were similar in morphology, marker expression, and teratoma formation. However there were significant differences (P < 0.05) between the differentiated cells formed by the different cell lines in levels of expression of tissue-specific markers such as renin, kallikrein, Glut-2, ${\beta}-$ and ${\delta}-globin$, albumin, and ${\alpha}1-antitrypsin$ (${\alpha}1-AT$). The hES cell lines also differed in proliferative activity. Our observations should be useful in basic and clinical hES cell research.