• Title/Summary/Keyword: 동시배양

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Algal Growth Potential (AGP) Assay Using Heterosigma akashiwo(Raphidophyceae) in Pukman Bay, Korea (Heterosigma akashiwo(Raphidophyceae)을 이용한 북만의 조류성장잠재력 시험)

  • Kim Mu-Chan
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.12 no.2 s.25
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    • pp.81-87
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    • 2006
  • Algal growth potential(AGP) assay using Heterosigma akashiwo was conducted in Pukman Bay. The effects of nutrients and microorganisms on the growth of H. akashiwo were specifically evaluated by the algal bioassay method. The different types of growth response of H. akashiwo to the addition of nutrients, and the co-incubation with microorganisms were clearly observed. Before H. akashiwo red tide occurrence, the growth of H. akashiwo was significantly stimulated by addition of nitrate of $50{\mu}M$ with phosphate of $5{\mu}M$. The addition of single phosphate had no clear effect on the growth of H. akashiwo. And the co-incubation with microorganisms had no clear effect on the growth of H. akashiwo. This result indicates that nitrate potentially limited the growth of H. akashiwo before red tide occurrence. However, during a bloom of H. akashiwo, the growth was significantly stimulated by addition of either nitrate of $50{\mu}M$ or phosphate of $5{\mu}M$. The addition of trace metals and vitamin $B_{12}$ had no clear effect on the growth of H. akashiwo in the period. This result indicates that both nitrate and phosphate potentially limited the growth of H. akashiwo during the bloom. On the other hand, during the termination period of H. akashiwo bloom, the growth of H. akashiwo was slightly stimulated by addition of phosphate and nitrate. But the growth of H. akashiwo was significantly enervated by the co-incubation with microorganisms. This result indicates that microorganisms potentially limited the growth of H. akashiwo in the period of bloom termination.

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Maturationof Hamster's Follicular Ova in Culture (Hamster 의 노포난자의 배양에 의한 성렬)

  • 조완규
    • The Korean Journal of Zoology
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    • v.10 no.1
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    • pp.1-9
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    • 1967
  • 본 실험은 4 일간의 성주기가 뚜렷한 성체인 golden hamster로부터 미성숙인 난자를 적출하여 화학적배양액내에서 그의 성숙을 유도시키고 성주기에 따른 미성숙 난자의 성렬율을 관찰하는 것을 주요 목적으로 하여 행하여졌다. 한 개의 난소로부터 5-6개의 미성숙 난자를 적출하여 5% bovine serum albumin이 섞인 TC Medium 199에 넣어서$CO_2$-incubator를 이용하여 6 시간내지 24시간 동안 37$^{\circ}C$를 유지해가며, 배양액이 직접 공기와접촉하는 것을 막기 위해 유동성인 paraffin oil로 배양액을 덮고서 배양을 완수했다. 실험의 결과를 다음과 같이 완수했다. 1. Hamster의 난자의 성숙분렬을 유발시키는데 가장 적합한 배양액은 BMOC, Eagle's Medium , Waymouth's Medium 그리고 TC Medium 199의 네가지 가운데 TC Medium199이었다. 즉 이 배양액을 이용했을 때 난자가 높은 성렬도를 보여주었다. 2. 5% bovine serum alumn을 TC Medium 199에 섞어주었을 때 미성숙 난자가 성순분렬을 유기하는 율이 가장 높았다. 3. 발정기에 있는 난소로부터 얻은 난자가 가장 현저하게 높은 율로 성숙분렬을 보여주었다. 반대로 발정후기의 난자는 배양 시작 후 단시간 내에 대부분이 퇴화하였으며 발정기에 가까워 갈수록 퇴화율이 줄어들었다. 이것은 노포가 성숙분렬을 억제하는 물질을 생성하리라고 여겨지고 있긴 \ulcorner만, 동시에 이 노포는 또 한편 난자의 배란뒤에까지도 생명력을 유지할수 있는 능력을 발정기에 이르기까지의 기간동안 난자에게 부여하며, 난자는 이 능력을 배란전까지 축적하게 되며 이 때문에 노포로부터 유리되어 나온 난자가 장기간 그 생명력을 유지해나가는 것이라고 추정된다. 4. Paraffin oil 로 공기를 차단하여서 배양한 난자나 혹은 watch glass를 이용하여 공기와 접촉시킨 난자에서나 모든 비슷한 율로 성숙분렬을 보여주었다. 이로 보아 조작이 까다로운 paraffin oil을 이용하는 방법보다는 손쉽게 배양할 수 있는 watch glass 의 방법이 오히려 유용하다고 할 수 있다.

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Changes of Fatty Acid composition During Dispecific culture of Scrippsiella trochoidea a Dinoflagellate and Pseudomonas spp. marine Bacteria (적조와편모조 Scrippsiella trochoidea와 해양세균 Pseudomonas spp.의 동시배양 시 지반산 조성의 변화)

  • 임월애;김학균
    • 한국해양학회지
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    • v.28 no.3
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    • pp.186-191
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    • 1993
  • Scrippsiella trochoidea is a dinoflagellate responsible for red tide in early spring in southern coastal water. Marine bacteria appear to exert critical roles on the development and decay of phytoplankton bloom in marine ecosystem. It is likely that marine bacteria, Pseudomonas spp., share some metabolic processes with S. trochoidea. To investigate interactions between S. trochoidea and Pseudomonas spp. directly, cysts of S. trochoidea isolated from the bottom mud in Masan Bay have been germinated and cultured. From the S. trochoidea cultured medium, we have isolated Pseudomonas spp., a dominant and cultured. From the S. trochoidea cultured medium, we have isolated Pseudomonas spp., a dominant species. Both of Pseudomonas spp. and S trochoidea have been simultaneously inoculated into the sterilized sea water and cultured to examine the change of fatty acids. The major fatty acids that showed increases in composition during the dispecific culture were $C_{18:0/},{\;}C_{20:5}{\;}and{\;}C_{22:5}$ in S. trochoidea, and in Pseudomonas spp. Especially, $C_{20:5}{\;}and{\;}C_{18:0}$ were increased in S. trochoidea but decreased in Pseudomonas spp. These results strongly suggest that two species share some processes in their fatty acid metabolism.

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Changes of Major Components and Microorganisms during the Fermentation of Korean Ordinary Kochujang (한국재래식(韓國在來式)고추장숙성중(熟成中)의 주요성분(主要成分) 및 미생물(微生物)의 변화(變化))

  • Ahn, Cheol-Woo;Sung, Nack-Kie
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.16 no.3
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    • pp.35-39
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    • 1987
  • The present study was attempted to obtain the basic data concerning a reasonable preparing method and the optimum fermentation conditions of Kochujang (Red pepper paste). To establish the standard qualify of Kochujang, changes of the chemical composition and the numbers of bacteria and feasts in Kochujang during fermentation were observed. Moisture, salts and crude ash contents of Kochujang were not changed significantly during fermentation. Titrable acidity and amino nitrogen gradually increased with the time-passed, whereas crude fat gradually decreased with the time-elapsed. And reducing sugar and total nitrogen increased until 40 days, but slightly decreased after this period. The numbers of bacteria and yeasts in the ingrients for the preparation of Kochujang were $3.9{\times}10^7/g$, $1.5{\times}10^3/g$ in red pepper powder, $7.6{\times}10^4/g$, $2.8{\times}10^2/g$ in salts. respectively, but those of sugar and malt were not more than 100/g. Microbial counts in Kochujang during fermentation increased until 40 days, but those are gradually decreased after that.

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A Study on Degradation Rate of Carbofuran in Sterilized Soil and Sulfate-added Paddy Soil (Carbofuran의 담수토양중(湛水土壤中) 분해(分解)에 대한 토양(土壤)의 멸균(滅菌)과 황산염(黃酸鹽) 첨가(添加)의 영향(影響)에 관(關)한 연구(硏究))

  • Tu, Ock-Ju
    • Korean Journal of Environmental Agriculture
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    • v.17 no.1
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    • pp.11-15
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    • 1998
  • Carbofuran was incubated for four weeks in five types of paddy soil samples at $25^{\circ}C$. The soil samples prepared in the study were as follows : control soil, sterilized soil, 10% cellulose added soil, 10% cellulose and 1% ferrous sulfate added soil, and 10% cellulose and 1% magnesium sulfate added soil. The degradation rate of carbofuran was significantly decreased(p<0.05) in sterilized soil.The degradation rate of carbofuran was significantly decreased by addition of cellulose(p<0.05) in femous sulfate added soil and magnesium sulfate added soil(p<0.01).

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Multiplex Reverse Transcription-PCR for Simultaneous Detection of Reovirus, Bovine Viral Diarrhea Virus, and Bovine Parainfluenza Virus during the Manufacture of Cell Culture-derived Biopharmaceuticals (세포배양 유래 생물의약품 제조공정에서 Reovirus, Bovine Viral Diarrhea Virus, Bovine Parainfluenza Virus 동시 검출을 위한 Multiplex Reverse Transcription-PCR)

  • Oh, Seon Hwan;Bae, Jung Eun;Kim, In Seop
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.339-347
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    • 2012
  • Viral safety is an important prerequisite for clinical preparations of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacturing process. In particular, Chinese hamster ovary (CHO) cells are highly susceptible to several RNA viruses including reovirus (Reo), bovine viral diarrhea virus (BVDV), and bovine parainfluenza virus (BPIV) and there have been reports of such viral contaminations. Therefore, viral detection during the CHO cell process is necessary to ensure the safety of biopharmaceuticals against viruses. In this study, a multiplex reverse transcription (RT)-PCR assay was developed and subsequently evaluated for its effectiveness as a means to simultaneously detect Reo, BVDV, and BPIV during the manufacture of cell culture-derived biopharmaceuticals. Specific primers for Reo, BVDV, and BPIV were selected, and a multiplex RT-PCR was optimized. The sensitivity of the assay for simultaneous amplification of all viral target RNAs was $7.76{\times}10^2\;TCID_{50}/ml$ for Reo, $7.44{\times}10^1\;TCID_{50}/ml$ for BVDV, and $6.75{\times}10^1\;TCID_{50}/ml$ for BPIV. The multiplex RT-PCR was proven to be very specific to Reo, BVDV, and BPIV and was subsequently applied to the validation of CHO cells artificially infected with each virus. It could detect each viral RNA from CHO cells as well as culture supernatants. Therefore, it was concluded that the multiplex RT-PCR assay can be applied to detection of the adventitious viruses during the manufacture of cell culture-derived biopharmaceuticals.

돼지 난소의 황체세포의 체외배양시 TGF-${\beta}$1의 분비

  • 최재혁;김봉기;정학재;박진기;우제현;양병철;장유민;박수봉;성환후
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.89-89
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    • 2003
  • 본 연구는 난소의 황체를 체외배양시 TGF-$\beta$1의 황체내 발현을 조사하기 위해 수행되었다. 돼지 황체는 축산기술연구소에서 사육중인 돼지(체중 145$\pm$kg) 12두로부터 발정을 유도시켜 배란 후 약 48시간째 도축하여 난소를 회수하였다. 회수된 난소로부터 황체를 분리하여 세절한 후 0.25% collagenase용액(0.025mg DNase, 50mM EDTA, 50mM Dithio-threitol)으로 37$^{\circ}C$의 진탕 수조에서 30분간 배양하여 황체세포를 분리 회수하였다. 회수된 황체세포는 D-MEM용액(GIBCO, 10% FCS와 antibiotics 첨가)으로 2회 세척하여 1$\times$$10^{6}$live cell/$m\ell$이 되도록 희석하여 24 well culture plate(Corning, New Tork 14831)에 분주하여 $CO_2$ 배양기($CO_2$: 5%)에서 24시간 간격으로 2회 배양액을 교환해 48시간 동안 배양하였다. 배양된 황체 세포는 immunocytochemistry 방법으로 TGF-$\beta$1의 발현을 관찰함과 동시에 황체조직도 같은 방법을 사용하여 TGF-$\beta$1 의 발현 유ㆍ무을 관찰하였다. 그 결과 황체세포 그리고 황체 조직 뚜렷한 TGF$\beta$1의 발현을 확인할 수 있었다. 이 결과로서 TGF$\beta$1은 황체기능을 유지하는데 하나의 인자로서 작용하며 다른 인자들과의 상호작용을 시사하고 있다.

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Gene Manipulation of Pin 2(Proteinase Inhibitor II) to the Cottonwood Leaf Beetle(Coleoptera : Chrysomelidae) in Transgenic Poplar(Populus deltodies × P. nigra) (형질전환(形質轉換)된 포플러의 딱정벌레에 대한 저항성(抵抗性) 유전자(遺傳子)(Proteinase Inhibitor II) 발현(發現))

  • Kang, Hoduck
    • Journal of Korean Society of Forest Science
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    • v.86 no.4
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    • pp.407-414
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    • 1997
  • The resistance of a non-transgenic poplar clone, 'Ogy' and three transgenic poplar lines to the cottonwood leaf beetle, Chrysomela scripta F., was evaluated by in vitro feeding. The lines were transformed with neomycin phosphotransferase II(NPT II) as a selectable marker, proteinase inhibitor II(pin2) as a resistance gene, and CaMV 35S as a promoter. An efficient method of sterilizing the beetle eggs and introducing them into plant tissue cultures was developed. The resistance of the transgenic lines was investigated in terms of effects tin leaf area consumed, insect weight, insect developmental stages, and plantlet root dry weight after feeding. Also, leaf area consumed was examined by leaf age as measured through leaf plastochron index(LPI). The leaf area consumed and insect weight were highly significant between transformants and control, and insect development in vitro was significant among the transgenic lines. Larval infestation was the most severe around LPI 4 to 5 which were young leaves. The system provided a quick, highly controlled method to screen developing transgenic plantlets directly.

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Effect of $Zn^{2+}$ and Ferulic Acid on Laccase and Manganese Peroxidase Production by Funalia trogii (Funalia trogii에 의한 Laccase와 Manganese Peroxidase의 생산시 $Zn^{2+}$ 및 Ferulic Acid가 미치는 영향)

  • Park, Chul-Hwan;Han, Eun-Jung;Lee, Byung-Hwan;Lee, Jin-Won;Kim, Sang-Yong
    • KSBB Journal
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    • v.21 no.2
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    • pp.85-89
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    • 2006
  • Typical property of the white-rot fungi is their ability to degrade lignin and other aromatic compounds with non-specific extracellular enzyme. In this work, the modification of the strain(Funalia trogii ATCC 200800) and the culture condition was performed to enhance enzyme productivity. Single cell was separated by the protoplasts formation and several putative laccase and manganese peroxidase inducers were tested. By adopting the modified strain, enzyme productivity increased comparing with that of the original strain. Extracellular enzyme formation was highly stimulated by the addition of copper and various aromatic compounds in the glucose-based culture medium.

Remediation of Soil Contaminated by Chlorinated Ethylene Using Combined Application of Two Different Dechlorinating Microbial Cultures and Iron Powder (두 종류의 탈염소화미생물 배양액과 철분 첨가에 의한 염화에틸렌 오염토양 복원)

  • Lee, Tae-Ho;Kim, Hyeong-Seok
    • Journal of the Korea Organic Resources Recycling Association
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    • v.11 no.2
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    • pp.55-65
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    • 2003
  • The combined effect of bioaugmentation of dechlorinating bacterial cultures and addition of iron powder($Fe^0$ on reductive dechlorination of tetrachloroethylene(PCE) and other chlorinated ethylenes in a artificially contaminated soil slurry(60micromoles PCE/kg soil). Two different anaerobic bacterial cultures, a pure bacterial culture of Desulfitobacterium sp. strain Y-51 capable of dechlorinating PCE to cis-1,2-dechloroethylene(cis-DCE) and the other enrichment culture PE-1 capable of dechlorinating PCE completely to ethylene, were used for the bioaugmentation test. Both treatments introduced with the strain Y-51 and PE-1 culture (3mg dry cell weight/kg soil) showed conversion of PCE to cis-DCE within 40days. The treatments added with $Fe^0$(0.1-1.0%) alone to the soil slurry resulted in extended PCE dechlorination to ethylene and ethane and the dechlorination rate depended on the amount of $Fe^0$ added. The combined use of the bacterial cultures with $Fe^0$(0.1-1.0%)) showed the higher PCE dechlorination rate than the separated application and the pattern of PCE dechlorination and end-product formation was different from those of the separated application. When 0.1% of $Fe^0$ was added with the cultures, the treatments with the strain Y-51 and $Fe^0$ resulted in cis-DCE accumulation from PCE dechlorination, but the treatment with the enrichment culture and $Fe^0$ showed the more extended dechlorination via cis-DCE. These results suggested that the combined application of and the bactrial culture, specially the complete dechlorinating enrichment culture, is practically effective for bioremediation of PCE contaminated soil.

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