• 제목/요약/키워드: agar overlay

검색결과 26건 처리시간 0.02초

구연산 생성 Candida lipolytica의 원형질체 융합 (Intraspecific Protoplast Fusion of Citric Acid Producer, Candida lipolytica)

  • 성낙계;심기환;전효곤;강신권;박석규
    • 한국미생물·생명공학회지
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    • 제13권4호
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    • pp.391-395
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    • 1985
  • 내산성이 우수하고 citric acid와 SCP를 많이 생산할 수 있는 효모균주를 육종하기 위하여 구연산 생성효모인 Candide lipolytica의 종내 원형 질체융합 조건을 검토하였다. 배양기간과 효소처리조건을 최적화함에 의해 98%의 protoplast가 형성되었다. 3 % agar와 30mM $CaCl_2$를 함유한 재생용최소배지에 동일배지의 중층에 의해 약 20-30%의 protoplast가 재생되었다. 2개의 영양요구성이 상보적인 영양요구성원형질체, L-14($lys^-$)와 T-24($try^-$)에 100mM $CaCl_2$를 함유하는 30% PEG 6000을 $30^{\circ}C$ 에서 20분간 처리하여 4-5${\pm}$$10^{-4}$의 융합빈도가 얻어졌다.

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Saccharomyces cerevisiae와 candida cariosilignicola사이의 세포융합에 관한 연구 (Protoplast fusion between saccharomyces cerevisiae and candida cariosilignicola)

  • 이재동;임하선
    • 미생물학회지
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    • 제26권1호
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    • pp.37-43
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    • 1988
  • This research was focused on investigation of the condition for protoplast formation and regeneration of protoplast fusion between Saccharomyces cerevisiae which has fermentation ability and Candida cariosilignicola which can grow at high temperature and utilize methanol. The results obtained were as follows; The highest production was collected in exponential growth phase. Ninety-nine% protoplast formation of C. cariosilignicola was obtained in glycin-NaOH buffer (pH10.0) containing Zymolyase 0.5mg/ml at $35^{\circ}C$ for 1hr incubation. The highest regeneration was produced when protoplast wuwpension containing 0.5% soft agar in buffered 50mM $CaCl_{2}$ was poured as a soft overlay onto 2% agar plates. Equal amuont of protoplast suspension of two strains was mixed and centrifuged. The subsequent pellet was added to 2ml of 35% polyethylene glycol (MW 4,000) containing 50mM $CaCl_{2}$, and incubated at $30^{\circ}C$ for 10min. Then 0.1ml of the suspension of aggregated protoplast was immediately covered with minimal medium and incubated at $40^{\circ}C$ for 5-7 days. As results, $SC_{1}$, $SC_{2}$, and $SC_{3}$ fusants were obtained. The physiological characteristics of fusants produced by protoplast fusion were; $SC_{1}$, and $SC_{2}$ utilized maltose, galactose, methanol, potassium nitrate. $SC_{3}$ utilized all the above materials except galactose.

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Cloning. Sequencing and Characterization of the Novel Penicillin G Acylase Gene from the Soil-isolated Leclercla adecarboxylata

  • Jun , Sang-O.;Lim, Ho-S.;Kim, Geun-Y.;Lee, Eung-S.;Lee, Mann-H.
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.331.3-332
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    • 2002
  • A novel penicillin G acylase (PGA)-producing bacterial strain was isolated from soil by using the Serratia marcescens overlay technique. The isolated strain was identified as Leclercia adecarboxylata based on the analyses of the biochemical characteristics (API 20E). the cellular fatty acid profile. and the 16S rDNA sequences. The gene encoding the PGA (pac gene) was cloned into the pHSG399 vector and the recombinant E. coliHB101 clones harboring the pac gene were isolated on agar plates containing phenylacetyl-L -leucine and penicillin G. (omitted)

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재생방법에 따른 교정용 브라켓의 세포독성에 관한 실험적 연구 (AN EXPERIMENTAL STUDY ON THE CYTOTOXICITY OF RECYCLED BRACKETS)

  • 임용규;양원식
    • 대한치과교정학회지
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    • 제23권2호
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    • pp.147-163
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    • 1993
  • The purpose of this stuy was to evaluated the cytotoxicity of brackets which were recycled thermally or chemically. New brackets and used brackets which had been in mouth for at least 2years were used as samples and human gingival cell culture and agar overlay technique was used to evaluate the cytotoxicity. From the experiment the following results were obtained : 1. New brackets in the as received state showed mild cytotoxicity. 2. Thermally recycled brackets except the used bracket not electropolished showed moderate cytotoxicity and among them new brackets showed greater cytotoxicity than used ones. 3. Used brackets which were thermally recycled without electropolishing showed mild cytotoxicity. 4. Among thermally recycled brackets, electropolished brackets showed greater cytotoxicity than not electro-polished ones. 5. Chemically recycled brackets showed moderate cytotoxicity, and among them, new brackets appeared to be more cytotoxic than used ones.

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교정용 호선의 세포독성에 관한 실험적 연구 (An experimental study on the cytotoxicity of orthodontic wires)

  • 임용규;양원식
    • 대한치과교정학회지
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    • 제26권5호
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    • pp.591-599
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    • 1996
  • 본 연구는 널리 이용되고 있는 두 종류의 교정용 호선에 다양한 처리를 가한 후 호선의 세포독성을 비교, 평가하고자 시행하였다. 018x025 inch 굵기의 stainless steel 호선과 Co-Cr 호선을 실험 재료로 선택하여 stainless steel 호선을 A 호선, Co-Cr호선을 B 호선이라 칭하였으며 각각의 호선을 다시 가해진 처리에 따라 4군으로 나누었다. A-1군과 B-1군은 제작된 상태 그대로의 호선을 이용하였으며 A-2, B-2군은 기기를 이용하여 $850^{\circ}\;F$에서 4분간 열처리하였다. A-3, B-3군은 같은 방법으로 열처리한 후 표면의 불순물을 제거하기 위해 전해연마를 시행하였고 A-4, B-4군은 소량의 은납(Ag-solder)을 납착(soldering) 하였다. 사람의 치은 섬유아세포를 배양하고 agar overlay법을 이용하여 각군의 호선의 세포독성을 검사하였으며 세포독성을 반응지수(탈색지수/용해지수)로 평가하여 다음의 결론을 얻었다. 1. stainless steel 호선과 Co-Cr 호선 모두 제작된 그대로의 상태에서는 세포독성을 나타내지 않았다. 2. 두 호선에 대한 열처리나 전해연마는 호선의 세포독성에 영향을 미치지 못하였다. 3. 은납이 납착된 stainless steel호선은 은납이 납착된 Co-Cr호선에 비해 더 넓은 범위의 탈색을 나타냈으나 탈색지수와 세포독성(반응지수)에서는 차이를 보이지 않았다. 4. 은납이 납착된 호선은 두 호선 모두에서 중증도의 세포독성을 나타냈다.

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Prophylactic Uses of Probiotics as a Potential Alternative to Antimicrobials in Food Animals

  • Lee, Hyeon-Yong;Xu, Hua;Lee, Hak-Ju;Lim, Tae-Il;Choi, Young-Beom;Ko, Jeong-Rim;Ahn, Ju-Hee;Mustapha, Azlin
    • Food Science and Biotechnology
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    • 제17권1호
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    • pp.191-194
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    • 2008
  • The antagonistic activity of probiotic strains (Bifidobacterium animalis BB-12, Bifidobacterium bifidum A, Bifidobacterium longum B6, Lactobacillus acidophilus ADH, Lactobacillus paracasei ATCC 25598, and Lactobacillus rhamnosus GG) against nalidixic acid resistant ($NA^R$) Escherichia coli O157:H7 MF1847, E. coli O157:H7 H2439, E. coli O157:H7 ATCC 43894, and E. coli O157:H7 C7927 was investigated using the agar-overlay, well diffusion, and broth culture tests. L. paracasei ATCC 25598 was the most effective probiotic strain in terms of in vitro antagonistic activity against $NA^R$ E. coli O157:H7, followed by L. rhamnosus GG, B. longum B6, and L. acidophilus ADH. The use of selected probiotic strains could be an effective pre-harvest intervention strategy to reduce the risk of $NA^R$ E. coli O157:H7 by maintaining a balanced microflora in animals and might provide many potential benefits in lieu of using antimicrobials.

수종(數種) 근관소독제(根管消毒劑) 및 근관충전재(根管充塡材)의 세포독성(細胞毒性)에 관(關)한 실험적(實驗的) 연구(硏究) (Evaluation of the Cytotoxicity of Root Canal Disinfectants and Root Canal Sealers on L Cells in Vitro)

  • 정충모
    • Restorative Dentistry and Endodontics
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    • 제7권1호
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    • pp.85-99
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    • 1981
  • This study was to evaluate the cytotoxic effect of three root canal disinfectants (formocresol, camphorated phenol and eugenol) and ten root canal sealers(Cavitec, Hypo-cal, Vitapex, AH26, Canals, Mynol, $N_2$, $N_2$-Medical, Z. O. E. and Calvital) in vitro. The experiments were performed in four differrent modes. In the first and second experiment, the "long-distance" cytotoxicity of three root canal disinfectants were tested on L cells. In the third exeriment, ten root canal sealers were tested for cytotoxicity by means of the tissue culture-agar overlay method immediately, 4 and 24 hours after the experiment. In the fourth experiment, the study with radioactively labeled L cells were employed to determine the relative cytotoxicity of ten root canal sealers. The results were as follows; 1. Every vapors from disinfectants showed more or less cytotoxicity. Of the three disinfectats, formocresol appeared to be the highest cytotoxic effect and camphorated phenol was the lowest. 2. Root canal sealers tested in tbis study showed cytotoxicity at every stage of time intervals. 3. The highest cytotoxic effect was freshly mixed $N_2$ meaical and $N_2$ also revealed the highest cytotoxic effect after 4 or 24 hours among these materials. Vitapex was found the lowest cytotoxic effect at all experimental stage. 4. Root canal sealers except N2 and Mynol showed cytotoxic effect were decreased cytotoxicity according to the time elapsed.

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수종 치과 교정용 밴드 시멘트의 세포독성에 관한 실험적 연구 (In vitro cytotoxicity of four kinds orthodontic band cements)

  • 이원철;박수병
    • 대한치과교정학회지
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    • 제34권4호
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    • pp.351-362
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    • 2004
  • 고정식 교정 치료시 교정용 밴드가 장착된 치아의 치주조직 손상은 접착된 치아에서 보다 자주 발생한다. 일반 치과치료에 사용되는 시멘트는 세포독성을 지닌 것으로 알려져 있으므로 치주조직과 직접 접촉하거나 구강 내에서 밴드시멘트의 성분들이 용출되는 교정용 밴드 시멘트는 치주조직의 손상을 일으키는 원인이 될 수 있다 그러므로 교정용 밴드 시멘트의 생체 친화성은 재료의 물성과 함께 중요하게 고려되어야 할 사항이다. 이 연구의 목적은 교정용 밴드 시멘트의 세포독성을 평가하는 것으로 현재 임상에서 널리 사용되고 있는zinc phosphate cement (ZPC), glass ionomer (GI), resin modified glass ionomer (RMGI), compomer의 생체 친화성을 평가하기 위해 human gingiva fibroblast를 배양하고 이 세포에 대한 세포수 산정 및 세포 형태관찰, MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, 한천중층시험 등의 세포독성 검사를 시행하여 다음과 같은 결과를 얻었다. 1. 세포수 산정 및 형태 관찰 시험에서 Gl>ZPC, RMGI, RMG124, GI24>compomer24, ZPC24, compomer 순으로 세포독성이 높게 나타났다. 2. MTT assay 에서는 GI>ZPC, RMGI>GI24>ZPC24, compomer, RMGI24, compomer24 순으로 세포독성이 높게 나타났다. 3. 한천중층시험에서 는 GI>GI24, ZPC, ZPC24, RMGI>RMG124, compomer, compomer24 순으로 세포독성 이 높게 나타났다.

Pseudomonas syringae pv. tabaci Phage의 분리 및 특성에 관한 연구 (Studies on the Isolation and Characterization of the Pseudomonas syringae pv. tabaci Phage)

  • 전홍기;김태인;유진삼;백형석
    • 미생물학회지
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    • 제32권1호
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    • pp.60-64
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    • 1994
  • 자연계로부터 식물병원균인 Pseudomonas syringae pv. Tabaci에 감염하는 bacteriophage 를 분리하였다. 이 phage의 안정성을 조사한 결과 중성 부근 pH에서 가장 안정하였고 50${\circ}C$ 이상에서는 안정성이 급격히 감소하였으며 흡착시간 별로는 10분까지는 빠른 흡착율을 보이다가 그 후부터는 서서히 감소하였다. 또한 흡착에는 금속이온을 필요로 하였으며 흡착온도는 20${\circ}C$에서 가장 높게 나타났고 20~40분에서 가장 높게 나타났다. 또한 배양온도에 따라 plaque 양상이 달랐는데 10${\circ}C$에서는 clear plaque를, 20, 30${\circ}C$에서는 turbid plaque를 형성하였다. 20${\circ}C$에서 잠복기는 약 3시간이었고 평균 방출수는 200PFU/cell이었다. 유전물질로는 ds DNA를 가지고 있었고 크기는 30kb 정도이었다.

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Rhizopus oryzae와 Aspergillus oryzae의 속간 원형질체융합 (Intergeneric Protoplast Fusion between Rhizopus oryzae and Aspergillus oryzae)

  • Lee, Soo-Youn;Jung, Sung-Won;Kim, Seong-Han;Lee, Yung-Nok
    • 미생물학회지
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    • 제31권3호
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    • pp.218-223
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    • 1993
  • Conditions for the release and regeneration of protoplasts form Rhizopus oryzae and intergeneric protoplast fusion between Rhizopus oryzae and Aspergillus oryzae were studied. High yields of protoplast fusion between Rhizopus oryzae and Aspergillus oxyzae were studied. High yield of protoplasts from young germilings of R. oryzae were obtained by using lytic enzymes containing chitosanase (3 mg/ml), chitinase (3 mg/ml) and Novozym 234 (5 mg/ml). 0.5M glucose was used as the osmotic stabilizer and optimum pH of buffer was determined to be pH 7.5-8.0. Under these conditions, protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts regenerated on solid medium with a soft agar overlay. We have also carried out protoplasts fusion between R. oryzae and A. oryzae and have succeeded in obtaining three types of intergeneric fusants. In these experiments, 35% PEG-4000 and 10 mM CaCl$_{2}$ were used as fsogenic agents, and auxotrophic properties were used as a genetic marker to select fusants. Complementation frequency be protoplasts fusion of A. oxyzae and R. oryzae was 4.4% * 10$^{-5}$ . The fusant strains of the first type were prototrophs showing an Aspergillus type morphology with dark-yellow sporulation, those of the second type were also Apergillus type morphology but showed no sporulation. And the strains of the third type stopped growing when fusion products grown on regeneration minimal medium were transferred to fresh minimal medium. The formation of fusion products was observed by fluorescent vital stains for complementary labelling of protoplats from R. oryzae and A. oryzae. Rhodamine 6G and fluorescein diacetate wer useful complementary vital stains of Rhizopus and Aspergillus protoplasts for visualization of requency and type (dicell, multicell) of fusion.

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