• Title/Summary/Keyword: E.coil

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Cloning and Expression of K11 Phage RNA Polymerase (K11 RNA 중합효소의 Cloning 및 발현)

  • Lee, Sang-Soo
    • The Journal of Natural Sciences
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    • v.9 no.1
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    • pp.19-24
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    • 1997
  • Using the PCR(polymerase chain reaction method), gone 1 of phage K11 coding for K11 phage RNA polymerase has been cloned and expressed under the control of lac promoter. K11 phage RNA polymerase was conventionally purified through the DEAE-sephacel and Affigel blue column chromatographies. The 0.2-0.3 M $NH_4Cl$ fractions of DAEA-sephacel column chromatography showed K11 phage RNA polymerase activity and further purification with Affigel blue column chromatography showed nearly single protein band on SDS-polyacryl amide gel. K11 phage RNA polymerase, which is one of the T7 group phage RNA polymerase (E. coil phage T7, T3 and Salmonella tyhimurium phage SP6 RNA polymerase), shares high degrees of homology with the other T7 group phage RNA polymerase. Previously we constructed T7 and SP6 promoter variants and revealed promoter specificity of T7 and SP6 RNA polymerase (Lee and Kang, 1993). To investigate the promoter specificity of K11 RNA polymerase in vitro K11 promoter activity was measured with SP6 promoter variants. The SP6 promoter variant share highest degrees of sequence homology with K11 promoter sequence show strongest promoter activity.

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Effects of Some Metabolic Inhibitors on Phototactic Movement in Cyanobacterium Synechosystis sp. PCC 6803 PTX (람세균 Synechocystis sp. PCC 6803 PTX의 주광성 운동에 미치는 몇가지 대사 억제제의 효과)

  • 박영총
    • Journal of Plant Biology
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    • v.38 no.1
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    • pp.87-93
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    • 1995
  • For understanding physiological nature of phototaxis in Synechocystis sp. PCC 6803 PTX(S. 6803 PTX), we examined the effects of some metabolic inhibitors and cation ionophore on the phototactic movement. In the presence of DCMU, which blocks the photosynthetic electron transport just after photosystem II acceptor, there was no inhibitory effect on the phototaxis up to $100\;\mu\textrm{M}$. Instead, the respiratory electron chain inhibitor such as sodium azide dramatically impaired the phototaxis in S. 6803 PTX. These observations indicate that the phototaxis is linked not to photo-phosphorylation, but to respiratory phosphorylation. When the cells were treated with un couplers such as CCCP or DNP, which dissipate the electrochemical gradient of proton($\Delta\mu_{H}+$) across the cytoplasmic membrane, these chemicals did not affect phototaxis. In contrast, when cells were treated with DCCD or NBD which deprive cells of A TP but leave $\Delta\mu_{H}+$ intact across the membrane, the phototactic movement was severly reduced. These results imply that ATP production, not proton motive force, is involved in the phototactic movement in this organism as a driving motive force. The application of specific calcium ionophore A23187 strongly impaired positive phototaxis. Calcium fluxes should be engaged in the sensory trans-duction of phototactic orientation. Finally, when ethionine was supplimented to culture media, the photomovement of this organism was inhibited. This implies that methylation/demethylation mechanism controls the process of phototaxis in S. 6803 PTX like chemotaxis in E. coli and Salmonella typhimurium.murium.

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The effect of Coptidis rhizoma, Chinese Galls and Phytoncide in Malassezia furfur (황련(Coptidis rhizoma), 오배자(Chinese galls), 피톤치드 혼합물(Phytoncide)이 비듬균(Malassezia furfur)에 미치는 영향)

  • Kim, Joung-Hee;Kwack, Seung Jun;Cho, Yong-Kwon;Kim, Byung-Kwan;Kim, Jong Guk;Lee, Eunkyung;Cho, Hye-Kyung;Kim, Keuk-Jun
    • Journal of Life Science
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    • v.25 no.7
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    • pp.748-756
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    • 2015
  • The report explores the possibility that the single extract of natural substances and the mixture of single extracts of natural substances create a synergy effect to increase the antimicrobial activity. It also compares the previous researches to find out if the natural medicinal herbs’ extract has antimicrobial activity on dandruff causative organism, Malassezia furfur. For the experiment on the mixture of single extracts of natural substances, the results are like following: 1. Staphylococcus aures’s antibacterial activity is resistant to mixture of three natural substances. 2. Escherichia coil’s antibacterial activity is resistant to mixture with Coptidis rhizoma. 3. Candida albicans’ antifungal activity is resistant to mixture with Chinese galls, which means the different results may be expected when tested with each germ. 4. On the other hand, Malassezia furfur has no antifungal activity on the single extract of natural substances and shows weak antifungal activity, whose diameter is 3.20 mm when tested with the mixture of 50% of Coptidis rhizoma and 50% of Phytoncide. The result is totally different from the one on the same eumycetes, C. albicans. That is because M. furfur has more lipophilic chemicaled cell walls than C. albicans does and it also consists of lamella layer, inner plasma membrane and intermediate multiple layers.

The Rate and Morphology of Mycorrhizal Infection in the Wild Ginseng (Panax ginseng C.A. Meyer) Collected from Various Locations in Korea (한국내 각지에서 채집된 산삼의 균근(菌根) 형태와 감염율)

  • Lee, Kyu-Hwa;Lee, Kyung-Joon;Park, Hoon;Budi Sri Wilarso
    • Journal of Ginseng Research
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    • v.30 no.4
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    • pp.206-211
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    • 2006
  • To investigate mycorrhizal infection by arbuscular mycorrhizal fungi(AMF), samples of fine lateral roots were taken from the wild ginseng(Panax ginseng C.A. Meyer) naturally growing at various locations in Korea. Mycorrhiazal infections were studied by cleaning the root samples and staining fungal hyphae with frypan blue. Wild ginsengs for this study were graded by an appraisal committee consisting of 12 experts of Korea Mountain Ginseng Association. Following five quality groups were recognized: Heaven group(pure natural), Earth group (from seeding of wild ginseng), Man group(from seeding or seedlings of wild ginseng with slight environmental modification), unmarketable, and imported wild ginseng. Morphology of AMF was typical Paris-type which shows intracellular hyphal coils with rare vesicles and lack of arbuscules. Average infection rate of individual wild ginsengs was 58.3% and showed no differences among five quality groups. When portions of fine roots were quantified for mycorrhizal infection, 18.7% of the total length of the primary and secondary roots were infected by AMF. Wild ginsengs from Gyeonggi Province(84.2%), and from mountains lower than 1,200 meters above sea level(about 70%) showed higher infection rate, while the ginseng from Gyeongbuk Province(27.8%) had lower rate. Wild ginsengs at older age showed lower infection rates.

Anti-wrinkle Activity of $\beta$-carotene Extracted & Purified from Recombinant Escherichia coli (재조합 대장균으로부터 추출.정제된 베타-카로틴의 주름개선 활성)

  • Jo, Ji-Song;Ku, Bo-Mi;Kang, Sang-Soo;Lee, Jae-Ran;Kim, You-Geun;Lee, He;Kim, Sung-Bae;Kim, Seon-Won;Kim, Chang-Joon;Chung, In-Young
    • KSBB Journal
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    • v.23 no.6
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    • pp.513-518
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    • 2008
  • This paper described the extraction/purification of $\beta$-carotene from recombinant E.coli and evaluation of anti-wrinkle activity of purified $\beta$-carotene. No significant differences in extraction yields were observed when hexane or isobutyl acetate was used. However, extraction from wet-cell cake resulted in 2-fold higher amount of $\beta$-carotene than that from dry cells. Disruption of 5 g-wet cells by ultrasonic homogenizer, acetone dehydration, extraction with isobutyl acetate resulted in 36 mg of $\beta$-carotene corresponding to 61.2% of recovery. The formation and separation of $\beta$-carotene crystal improved the purity. 633 mg of $\beta$-carotene crystal with 93% purity was obtained from 223 g/L of wet-cell cake harvested from 2.5-L fed-batch culture broth. The cultures of normal human primary fibroblast were performed to investigate the effect of $\beta$-carotene on cytotoxicity as MTT assay and anti-wrinkle activity as collagen synthesis assays. $1.7{\mu}M$ of $\beta$-carotene was found to be optimal concentration at which 1.4-fold higher amount of collagen was synthesized than that in absence of $\beta$-carotene. This indicates that highly purified $\beta$-carotene can be obtained from recombinant E.coli by applying simple method with less toxic solvent and can be used in functional cosmetics as anti-wrinkle agent.

Heating Performance of Hot Water Supplying System in Greenhouse (온수배관을 이용한 온실의 난방성능)

  • Yoon, Yong-Cheol;Shin, Yik-Soo;Kim, Hyeon-Tae;Bae, Seoung-Beom;Choi, Jin-Sik;Suh, Won-Myung
    • Journal of Bio-Environment Control
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    • v.21 no.2
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    • pp.79-87
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    • 2012
  • This research was conducted to obtain basic data with regard to the heating performance that would be produced by installing an aluminum hot water pipe inside the greenhouse with the goal of reducing the heating energy in greenhouse. The research results are summarized as follows. The degree of difference in relation to the temperature by height within the greenhouse during the entire experiment was significant - within the range of 4.0~$7.0^{\circ}C$. The temperature difference between incoming and outgoing water was about $3.3^{\circ}C$ greater when FCU was activated compared to when it was not activated. Meanwhile, the amount of energy consumed increased about 36.2~40.1%. The amount of pyrexia per hour also increased by 44.6~52.0%. During the experiment period, circulated flux was within the range of 0.48~$0.49L{\cdot}s^{-1}$ while average fluid speed was 1.53~$1.56m{\cdot}s^{-1}$. The average temperature difference between incoming and outgoing water was 6.24~$11.50^{\circ}C$. The amount of heating value by each set temperature within the minimum outdoor temperature range of -14.0~$-0.6^{\circ}C$ was 135,930~307,150 kcal, and the range was within the 9,610~$19,630kcal{\cdot}h^{-1}$ per hour. This demonstrated that about 23~53% heating energy of the maximum heating load could be supplied. Total radiating value and amount of energy consumed were 2,548,306 kcal and 3,075.7 kWh, respectively. When heating takes place using oil, which is a fossil fuel, the total amount of light oil consumed was 281.6 L while the cost was 321,000 won. When the electricity cost for farms is applied, the total cost was about 110,730 won, which is about 33.5% of the cost required compared to oil consumption. The temperature at in the experiment area was about 8.3~$14.6^{\circ}C$ higher compared to that of the control area.