• Title/Summary/Keyword: 계대배양

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Change of Infection Properties of Subcultured Marine Birnavirus in Several Fish Cell Lines (어류 주화세포에서의 계대배양에 의한 해양버나바이러스의 감염특성의 변화)

  • Jung, Sung-Ju
    • Journal of fish pathology
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    • v.11 no.2
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    • pp.89-96
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    • 1998
  • Marine birnavirus (MABV) has wide host range in marine organisms. To clarify various infection properties of MABV in different host species, in vitro study was performed by subculture for 10 passages in several fish cell lines. In CHSE-214, RTG-2 and RSBK-2 cells, the virus produced high yield of virus. Typical CPE with high protein expression was observed in these cells. On the contrary, the virus grown in EPC, FHM and BF-2 cells exhibited no CPE appearance although virus protein was detected. In EPC and FHM cells, the virus titer increased in later passages. The plaque size was distinctly bigger in CHSE-214, RTG-2 and RSBK-2 cells than in other cell lines. The nucleotide sequence of VP2/NS junction region on genome segment A exhibited one specific nucleotide change at 195. The different infection properties in several cell types performed in the present work might reflect in vivo MABV infection in various host species occurring in natural conditions.

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Proliferation of Gladiolus 'Topaz' Callus by Liquid Shaking Culture (액체진탕배양에 의한 글라디올러스 'Topaz' 캘러스의 증식)

  • 최정두;김규원
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.157-161
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    • 1999
  • This study was performed to enhance the proliferation rate of Gladiolus 'Topaz' callus. The callus was induced from the cermet tissue explants on MS solid medium with 10 mg/L 2,4-D. In the case of liquid shaking culture, proliferation of the callus was effective in MS medium with 0.05 mg/L 2,4-D at 2$0^{\circ}C$ under 16 hours daylength and in a 100 mL Erlenmeyer flask containing 20 mL of the liquid medium and at 75 rpm in rotation speed of the horizontal shaking culture. Furthermore the callus was also able to be subcultured in the same liquid medium.

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Axenization of Entamoeba histolytica, a Korean strain YS-27 (이질아메바 한국분리주 YS-27의 무균 배양화)

  • Jang, Jae-Gyeong;Im, Gyeong-Il;So, Jin-Tak
    • Parasites, Hosts and Diseases
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    • v.33 no.4
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    • pp.387-390
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    • 1995
  • In the present communication, YS-27, a Korean strain of Entnnloeba histoIWtica is described for the isolation and establishment of axonic cultivation. 5. histoLvticc, designated as strain "YS-27" was isolated from the pus of a hepatic abscess obtained from a 72 you old inpatient of August 10, 1969. Specimens, were obtained by needle aspiration, inoculated immediately and weekly cultured in a modified diphasic medium at 37℃. Strain YS-27 had been maintained for more than 15 years by weekly subculture until February, 1985. These cultures were transferred to a monoxenic TTY-SB medium seeded with a trypanosomatid of the genus CyithidiG. Penicillin G, 2 to 10 H 103 International units and Streptomycin, 2 to 10 mg per 100 ml, were added to the cultures to eliminate the bacteria. After more than one year later, these two organisms were well maintained by transfer every 3 or 4 days until .January. 1986 at 37℃ in TTY-SB medium in the absence of other microorganisms. These monoxenic cultures were then transferred to TYI-S-33 medium. Strain YS-27 alone had not been growing at the time of transfer, but when overlaid with Crithinia at intervals of 3 to 4 days, strain YS-27 propagated well. The Clthidio died out several weeks later after several passages. Beginning in April, 1986, strain YS-27. was successfully established in axonic culture in TYI-S-33 medium and has been maintained in continuous culture and multiplied well to present.

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Studies on Proper Medium for Somatic Embryogenesis in Suspension Culture of Rehmania glutinosa and Encapsulation of Somatic Embryos (지황의 현탁배양에서 체세포배 형성에 관여하는 요인분석과 체세포배의 Encapsulation)

  • Park, Ju-Hyun;Park, Sang-Un;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.2
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    • pp.100-106
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    • 1995
  • This study was conducted to find the factors affecting somatic embryogenesis in suspension culture of Rehmania glutinosa and investigate the possibility of artificial seed production by encapsulation of somatic embryos. Linsmeier-Skoog medium was appeared as proper for somatic embryogenesis. Sucrose with $3{\sim}5%$ as carbon sources was good for somatic embryogenesis, and both ammonium and nitrate nitrogen were necesary for normal somatic embryo production. BA with NAA or kinetin with NAA were better than the use of cytokinin alone for both somatic embryogenesis and numbers of somatic embryos. $AgNO_3$ as protectant for vitrification of seedlings in vitro culture had no harmful effect on somatic embryos. Sphericity of encapsulated seeds was good at 3% gel of sodium alginate but germination was better at 2.5% sodium alginate level. Artificial seeds were germinated and developed normal shoots and roots under in vitro condition.

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Improvement of Peroxidase Productivity by Optimization of Medium Composition and Cell Inoculum Size in Suspension Cultures of Sweet Potato (Ipomoea batatas) (고구마(Ipomoea batatase)현탁배양에서 배지조성 및 세포접종량의 적정화에 의한 Pemxidase생산성 향상)

  • 곽상수;김수경;정경희;유순희;박일현;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.91-97
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    • 1994
  • To improve the productivity of peroxidase (POD) of cell line SP-47 derived from cell suspension cultures of sweet potato (Ipomoea batatas (L) Lam.cv White Star), we optimized culture conditions including the composition and concentration of plant growth regulators and carbon source, and the cell inoculum size. When one g (fr wt) of cells was inoculated into 50 mL TL medium supplemented with l mg/L 2,4-D and 30g/L sucrose in 300 mL Erlenmeyer flask at 25$^{\circ}C$ in the dark (100rpm), the POD activity per g cell dry wt was maximized to be about 6,800 units after 25 days of subculture, which was about 30 times higher than that of intact roots of horseradish plants grown in the greenhouse, but the cell growth was maximum after 15 days of subculture. The protein content per g cell dry wt maintained almost plateau and after 25 days of subculture decreased as culture Proceeded further whereas the POD specific activity (unit/mg protein) was about two times higher after subculture and continuously increased from 12 days to the end of cultures (40 days). The POD isozyme patterns showed almost the same regardless of cell growth stage, but some acidic isozymes were slightly increased after 25 days of subculture. These results indicate that POD activity in suspension cultures of sweet potato is closely associated with cell growth and stresses derived from cell culture renditions and medium depletion. Due to its high POD activity the SPL47cell line seems to be suitable for the mass production of POD.

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Mass Production of Acanthopanax pedunculus through embryogenic cell culture and of madicinal row materials through bioreactor culture (참오갈피(Acanthopanax pedunculus)의 배형성 세포배양을 통한 식물체 대량번식 및 bio-reactor를 통한 약용원료 생산)

  • 정수진;윤의수;최용의
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.29-29
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    • 2002
  • 참오갈피(Acanthopanax pedunculus)는 국내 고유한 자생 종이며 현재 몇 그루 만이 존재하는 것으로 추정된다. 이 수종은 대체적으로 다른 오갈피 수종에 비해 잎 및 열매는 물론 식물 전체 부위가 다소 커서 경제성이 높다고 판단된다. 본 실험에서는 참오갈피의 세포배양 기술을 이용한 묘목 및 약용원료의 산업적 대량 생산 기술을 개발하고자 수행 하였다. 참오갈피 접합자배를 적출하여 무균적으로 배양한 다음 자엽단계로 성숙한 이들 접합자배를 1 mg/1 2,4-D가 첨가된 MS 배지에 배양하여 배형성 캘러스를 유도하였다. 배형성 세포의 유도율은 약 72%에 달했다. 유도된 배형성 캘러스는 2,4-D가 첨가된 같은 고체 및 액체 배지에서 계대배양하여 배형성 캘러스 및 세포를 유지하였다. 배형성 캘러스를 2,4-D가 첨가되지 않은 배지에 옮겨주면 이들 세포로부터 체세포배가 발생되었다. 약 2개월의 기간을 거쳐 자엽단계로 성숙된 체세포 배는 발아되지 않기 때문에 5 mg/1 GA$_3$가 첨가된 1/2 MS 배지에 옮겨 발아 및 식물체로 재생 시켰다. 약 7 cm 크기로 재생된 식물체를 인공토양에 옮겨 1달간 순화시키고 난 후 토양에 옮겼을 경우 87%가 생존하였다. 기내에서 유도된 참오갈피 식물체는 특별한 처리를 하여주지 않아도 뿌리로부터 배발생 세포를 형성 할 수 있었다. 따라서 이들 식물체로부터 배발생세포 유도는 세포주의 보존 및 증식에 효과적으로 이용될 수 있었다. 한편 참오갈피 세포는 5-10 리터 바이오리엑터 배양을 통하여 배형성세포, 자엽단계의 배, 유식물의 대량생산의 가능성을 확인하였다.

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Effect of Incubation Time, Concentration of Enzyme, and 2,4-D on Isolation and Callus Formation of Protoplast from Callus of Citrus junos (遊離시간 , 酵素處理 및 2,4-D 농도가 재래 유자(Citrus junos)의 캘러스由來 原形質體 遊離 및 培養에 미치는 영향)

  • 오성도;김영숙
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.335-339
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    • 1998
  • The factors affecting the isolation and culture of the protoplast of embryogenic callus, derived from immature ovule in Citrus junos, were examined. An incubation time in enzyme solution of 16 hrs was preferable for protoplast isolation. Efficient protoplast yields were obtained from the treatment of equal concentration of 0.7 M $\textrm{BH}_{3}$ to the enzyme solution containing 1.0% cellulase, 1.0% macerozyme and 0.2% pectolyase. Protoplast cultured in MT medium with 0.1 mg/L 2,4-D showed vigorous division and some of them formed callus. Induced callus was subcultured on solid MT medium but the callus showed very slow growth. The above results show the possibility to culture from protoplast fusion in Citrus genera.

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Histological Observation of Embryogenic and Non-embryogenic Callus in Long-term Subculture of Wild Viola (Viola patrinii DC.) (흰제비꽃 배양세포에 있어서 분화세포와 미분화세포 조직의 비교 관찰)

  • 정용모;손병구;이재헌;서정해;정정한;권오창
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.233-238
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    • 2000
  • To obtain a basic information of the development of Genus Viola, morphological and histological observation of in vitro calli and cells in Viola culture cells were investigated. There were two callus types obtained by long term subculture of wild viola (Viola partrinii DC. ) petiole callus. One was friable callus - soft and pale green in color and small cells in size, and the other was compact callus - compact and deep bluish green in color, large cells in size. In scanning electron microscopic observation, friable callus was composed of voculated cell around small. cell clump, while compact callus was composed of cells filled with protoplasm Somatic embryogenesis was observed from suspension culture of the compact callus.

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Effects of Donor Somatic Cell Conditions on In Vitro Development of Nuclear Transplanted Porcine Embryos (돼지 공여세포의 조건이 핵이식 수정란의 체외발달에 미치는 영향)

  • 홍승표;박준규;이명열;이지삼;정장용
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.213-221
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    • 2001
  • This study was conducted to examine in vitro developmental ability of porcine embryos after somatic cell nuclear transfer. The porcine ear fell was cultured in vitro for confluency in serum-starvation condition(TCM-199 + 0.5% FBS) far 3~6 days of cell confluency. The zona pellucida of IVM oocytes were partially drilled using laser system. Single somatic cell was individually transferred into enucleated oocytes. And the reconstructed embryos were electrically fused(single DC 1.9kv/cm, 30$\mu$ sec) with 0.3M mannitol. After electrofusion, embryos were activated(single AC 5v/mm, 5sec) and cultured in HCSU-23 medium containing 10% FBS at 39$^{\circ}C$, 5% $CO_2$ in air for 6 to 8 days. The fusion rate of donor cells was 45.6, 36.8 and 46.1% in 3~4, 5~6 days of serum starvation and non serum starvation(N-S), and were 52.7. 53.0 and 51.7% in 1~2. 5~6 and 13~14 passages of donor cell culture, respectively. No significant difference was found in the fusion rate of donor cells by the duration of serum starvation treatment or the number of donor cell passages. By the size of donor cells, however, the fusion rate was significantly higher(P<0.05) for reconstructed embryos derived from 25r $\mu$m $\geq$ site of donor cells (65.3%) than that of 25~30$\mu$ m(42.5%) or 30$\mu$ m(45.5%)$\leq$ cells. The cleavage rate was significantly (P<0.05) higher in 3~4 darts of serum starvation treatment(67.1%) than that in N-S (50.7%) or 5~6 days of starvation(57.1%). The activation rate by the size of donor cells in fused oocytes was 56.5, 68.8 and 58.5%, respectively, and was not significant.

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Callus induction and plant regeneration from in vitro cultured petiole of 3 Gerbera cultivars. (거베라 기내 엽병조직으로부터 Callus 유도 및 식물체 재분화)

  • 정용모;나애실;고은경;송혜정;김정부;권오창
    • Journal of Life Science
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    • v.14 no.5
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    • pp.855-858
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    • 2004
  • The experiment was conducted to investigate optimal condition for callus induction and plant regeneration for transformation system of gerbera. Callus induction was more effective in 'white day' then other two cultivar 'Songsongee' and 'Love Song' The optimized plant growth regulators concentration on callus induction, was MS basal medium with NAA 0.1 mg/L+ TDZ 0.5 mg/L. The optimized plant growth regulators concentration on plant regeneration, which was used MS basal medium was IAA 1.0 mg/L + BA 1.0 mg/L + Zeatin 0.1 mg/L. The optimized petiole age for more effective plant regeneration was 32 days petiole after in vitro subculture and MS basal medium strength was 1/2 MS strength.