• Title/Summary/Keyword: Lac

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Identification and Characterization of pH-Regulated Genes in Saccharomyces cerevisiae

  • Hong, Sung-Ki;Choi, Eui-Yul
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.327-333
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    • 1996
  • Yeast, like many other microbes, encounters large variations in ambient pH in their natural environments. Microorganisms capable of growing over a wide pH range require a versatile, efficient pH homeostatic mechanism protecting intracellular processes against extremes of pH. In several organisms, fusions to the bacterial lacZ gene have been extremely useful for the identification of genes expressed at different time during the life cycle or under different growth conditions. In this study, using the lacZ gene screening system, we surveyed a large number of yeast strains with lacZ insertion to identify genes regulated by pH. A yeast genomic library was constructed and inserted with lacZ by a shuttle mutagenesis procedure. The yeast transformants were individually picked up with a toothpick, replica-plated, and grown in alkaline pH medium. Among the 35,000 colonies screened, 10 candidate strains were identified initially by the $\beta$-gal assay. We finally confirmed two yeast strains carrying the genes whose expression are strictly dependent on pH of growth medium. One of the fusions showing a 10-fold induction in expression level in response to alkali pH was selected and further characterized. The pH-regulated gene was cloned by inverse PCR and a partial sequence of the gene was determined. Identification and characterization of the gene is currently under investigation.

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Genetic Responses to Metal ion in Aslmonella typhimurium (Salmonella typhimurium의 금속이온에 대한 유전적 반응)

  • Jung, Ju-Ri;Park, Kyeong-Ryang;Koh, Sang-Kyun;Park, Yong-Keun;Lee, In-Soo
    • Journal of Life Science
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    • v.8 no.2
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    • pp.216-225
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    • 1998
  • Metal ion-induced and it’s regulatory genes were screened in virulent salmonella typhimurium UK1 and tested cross-regulation with various stresses. Using the techniqud of P22-MudJ(Km, lacZ)-directed lacZ operon fusion, LF40 cuiA::MudJ and Lf153 cuiD::MudJ which were induced by copper were selected. cuia and cuiD were determined anaerobic coper inducible and copper tolerance response gene, respectively. Also cuiA and cuiD locus were determined at 81 and 8min, respectively, on salmonella Genetic Map. The two regulators were identified as cuaR, and cudR, which controls cuiA and cuiD, respectively. cuaR, and cudR appeared as negative regulators because the expression of cuiA-lac-Z and cuiD-lacZ were increased. Copper adapted UK1 showed high resistance to H$_{2}$O$_{2}$, but cuiD did not. The product of the cudR locus was responsible for decreasing the tolerance to copper and H$_{2}$O$_{2}$. Furthemore cuiA and cuiD locus were found to be part of a regulon under the control of a trans-acting regulators, rpoS, oxyR and relA. Therefore, the results suggest CTR participate with oxidative stress on Salmonella.

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raf Proto-oncogene is Involved in Ultraviolet Response in Drosophila

  • Ha, Hye-Yeong;Yu, Mi-Ae
    • Animal cells and systems
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    • v.1 no.4
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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.

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Cloning and Transcriptional Fusion with lacZ of a Gene (exo) Required for Exo-polysaccharide Synthesis in Rhizobium fredii USDA191 (Rhizobium fredii USDA191의 체외다당류 합성관련 유전자(exo)의 클로닝 및 lacZ와의 융합)

  • 정완석;고영환
    • Korean Journal of Microbiology
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    • v.31 no.1
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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.

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The Effects of Lactic Acid Bacteria on the Growth and Aflatoxin $B_1$ Production of Aspergillus parasiticus (유산균이 Aspergillus parasiticus의 성장과 Aflatoxin $B_1$ 생성에 미치는 영향)

  • Lee Kyung-Min;Kim Jong-Gyu
    • Journal of Environmental Health Sciences
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    • v.31 no.2 s.83
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    • pp.127-133
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    • 2005
  • This study was performed to investigate the competitive effect of Leuconostoc mesenteroides and Lactobacillus plantarum on the growth and aflatoxin $B_1$ production of Aspergillus parasiticus. Leu. mesenteroides and Lac. plantarum were grown with A. parasiticus in a modified APT broth at $28^{\circ}C$ for 9 days. It was observed that the growth of A. parasiticus got lower when the two bacteria were inoculated simultaneously than when the mold grew alone. The pH of mixed culture of Leu. mesenteroides and Lac. plantarum showed significantly lower value than the pH of the pure culture of A. parasiticus (p<0.05). The acidity of the mixed culture group significantly increased compared with the control group (A. parasiticus alone group) (p<0.05). The aflatoxin $B_1$ production was significantly decreased in the mixed culture group than in the A. parasiticus alone group (p<0.05). Leu. mesenteroides showed the more efficient effect than Lac. plantarum. These results indicate that the two lactic acid bacteria have inhibitory effect on the growth and aflatoxin $B_1$ production of Aspergillus parasiticus.

Device Optimization of N-Channel MOSFETs with Lateral Asymmetric Channel Doping Profiles

  • Baek, Ki-Ju;Kim, Jun-Kyu;Kim, Yeong-Seuk;Na, Kee-Yeol
    • Transactions on Electrical and Electronic Materials
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    • v.11 no.1
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    • pp.15-19
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    • 2010
  • In this paper, we discuss design considerations for an n-channel metal-oxide-semiconductor field-effect transistor (MOSFET) with a lateral asymmetric channel (LAC) doping profile. We employed a $0.35\;{\mu}m$ standard complementary MOSFET process for fabrication of the devices. The gates to the LAC doping overlap lengths were 0.5, 1.0, and $1.5\;{\mu}m$. The drain current ($I_{ON}$), transconductance ($g_m$), substrate current ($i_{SUB}$), drain to source leakage current ($i_{OFF}$), and channel-hot-electron (CHE) reliability characteristics were taken into account for optimum device design. The LAC devices with shorter overlap lengths demonstrated improved $I_{ON}$ and $g_m$ characteristics. On the other hand, the LAC devices with longer overlap lengths demonstrated improved CHE degradation and $I_{OFF}$ characteristics.

Production of Mn-peroxidase and Laccase from Lentinus edodes and Coriolus versicolor (표고 및 구름버섯으로부터 Mn-peroxidase와 Laccase의 생산(生産))

  • Bae, Hyeun-Jong;Han, Ok-Soo;Koh, Hong-Bum;Kim, Yoon-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.21 no.3
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    • pp.87-93
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    • 1993
  • This study was undertaken to investigate the characteristics and the productivities of lignin olytic enzymes: laccase (Lac) and Mn-dependent peroxidase (MnP) from Coriolus versicolor and Lentinus edodes respectively. Enzymes were isolated from cultural filterates and purified according to the standard methods. These enzymes showed one band in SDS-PAGE and their molecular weights were found 62,000 and 45,000 dalton respectively. Polyclonal antibodies against Lac and MnP were raised against mouse. In the ELISA (enzyme-linked immunosorbent assay), Lac and MnP-antiserum produced a strong positive reaction with Lac and MnP antigen($A_{405}$=2.50 and 3.53 respectively). The sera to negative (S/N) ratio was determined by the dividing the mean absorbance of antibodies by the corresponding diluted samples from normal mouse serum. The sera produced showed 2 times more positive reaction in S/N ratio than negative sera.

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SPINK1 promotes cell growth and metastasis of lung adenocarcinoma and acts as a novel prognostic biomarker

  • Xu, Liyun;Lu, Changchang;Huang, Yanyan;Zhou, Jihang;Wang, Xincheng;Liu, Chaowu;Chen, Jun;Le, Hanbo
    • BMB Reports
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    • v.51 no.12
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    • pp.648-653
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    • 2018
  • Serine protease inhibitor Kazal type 1 (SPINK1) plays a role in protecting the pancreas against premature activation of trypsinogen and is involved in cancer progression. SPINK1 promoted LAC cells growth, migration, and invasion. Mechanistically, we found that SPINK1 promoted LAC cells migration and invasion via up-regulating matrix metalloproteinase 12 (MMP12). We observed that SPINK1 expression was only up-regulated in lung adenocarcinoma (LAC) tissues, and was an independent prognostic factor for poor survival. Our results indicate that SPINK1 might be a potential biomarker for LAC that promotes progression by MMP12.

Direction for Improving Management Model based on Carrying Capacity for Managing natural resources in ecotourism area : Focused on Getz, LAC, VIM and VERP (생태관광지역 자연자원 관리를 위한 수용력 기반 관리모델 개선방향 : Getz, LAC, VIM, VERP 관리모델을 중심으로)

  • Park, Hong Chul;Oh, Choong Hyeon
    • Journal of Environmental Policy
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    • v.13 no.4
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    • pp.107-133
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    • 2014
  • The eco-tourism has caused a variety of problems between use and conservation. The situation requires introduction of scientific management techniques, such as controlling the number of visitors to the destination and identifying appropriate carrying capacity. Getz, LAC, VIM and VERP have been developed for this purpose and they are the management model based on carrying capacity that have been explored for introduction and application in Korea. However, it is necessary to modify and supplement the management model to suit the purpose of ecotourism characteristics and the status of ecotourism in Korea. Therefore, we identified the problems of the existing management model and presented three improvements. strengthen the interdependent linkages between carrying capacity and the management goals; improve the tourism opportunity distribution model; improve the phased management. It will help many ecotourism sites to effectively manage the impacts of visitors in Korea with a minimum amount of effort and costs.

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Migration Activity of Chicken Gonadal Primordial Germ Cells (gPGCs) and Post-transfer Localization of LacZ-transfected gPGCs in the Embryonic Gonads

  • Jeong, D.K.;Han, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.9
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    • pp.1227-1231
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    • 2002
  • A powerful tool for chicken transgenesis could be established by employing a germline chimera production through primordial germ cell transplantation. This study was conducted to examine whether foreign gene-transfected gonadal primordial germ cells (gPGCs) have a migration activity into the gonad after transfer to recipient embryos. In Experiment 1, gPGCs of Korean Ogol Chicken were retrieved from 5.5-day-old embryos and subsequently transferred to the dorsal aorta of 2.5-day-old White Leghorn embryos after being labeled with PKH26 fluorescent dye. To confirm migration activity after transplantation, recipient embryos were sacrificed and examined on 3 days after transfer. Sex determination was concomitantly undertaken to examine whether sex of recipient embryos could affect the migration activity of gPGCs. All of embryonic gonads examined showed positive signals with PKH26 fluorescence and W-chromosome specific band by polymerase chain reaction (PCR) was detected in male embryos when gPGCs with ZW chromosome were transferred to recipient embryos. In Experiment 2, retrieved gPGCs were transfected with LacZ gene-containing cytomegalovirus promoter ($pCMV{\beta}$) by electroporation and subsequently transferred to recipient embryos. LacZ gene expression was identified in the gonads of 6 or 10-day-old recipient embryos and hatched-chicks. A total of 20 embryos and 12 hatched-chicks were examined and 11 of them (10 embryos and one hatched chicken; 11/32=34.4%) expressed $\beta$-galactosidase, a marker substance of LacZ gene. The results of this study demonstrated that foreign gene-transfected gPGCs can migrate and settle down into the gonad after being transferred into the blood vessel of the recipient embryos. This established technique will contribute to developing a peer biotechnology for transgenic chicken.