• Title/Summary/Keyword: 계대 배양

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Effect of the Isolation Method of Mouse Inner Cell Mass, Types of Feeder Cells and Treatment Time of Mitomycin C on the Formation Rate of ICM Colony (생쥐 내세포괴의 분리방법과 지지세포의 종류와 Mitomycin C 처리 시간이 내세포괴 Colony 형성률에 미치는 영향)

  • Jang, Ho-Jin;Ko, Kyung-Rae;Kim, Mi-Kyung;Na, Yong-Jin;Lee, Kyu-Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.4
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    • pp.265-272
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    • 2006
  • Objective: This study was carried out to evaluate the effect of the isolation methods of inner cell mass from mouse blastocyst, types of feeder cells and treatment time of mitomycin C on the formation rate of ICM colony. Methods: The inner cells were isolated by conventional immunosurgery, partial trophoblast dissection with syringe needles and whole blastocyst co-culture method. Commercially available STO and primary cultured mouse embryonic fibroblast (pMEF) feeder cells were used, and mitomycin C was treated for 1, 2 or 3 hours, respectively. The formation rate of ICM colony was observed after isolation of ICM and culture of ICM on the feeder cells for 7 days. Result: The ICM colony formation rate on STO were significantly higher in partial trophoblast dissection group (58%) than that in immunosurgery (12%) or whole blastocyst culture (16%) group (p<0.05). The formation rate on pMEF feeder layer was higher in partial trophoblast dissection (88%) and whole blastocyst culture (82%) group than that in immunosurgery (16%) group (p<0.05). When mitomycin C treated to pMEF for 2 hours, the formation rate of 88% was significantly higher than those of other conditions. Conclusion: Above results showed that the efficient isolation method of ICM from blastocyst was the partial trophoblast dissection and the appropriate treatment time of mitomycin C was 2 hours. However, the subculture of ICM colony and characterization of stem cells should be carried out to confirm the efficacy of the partial trophoblast dissection method.

Plant Regeneration and Protein Analysis from Cadmium Resistant Callus of Tobacco (Nicotiana tabacum cv. BY4) (담배 (Nicotiana tabacum cv. BY4)카드뮴 저항성 캘러스로부터 식물체 재생과 단백질 분석)

  • 오승철;소웅영;조덕이;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.7-13
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    • 2001
  • Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

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Development of Pichia stipitis Co-fermenting Cellobiose and Xylose Through Adaptive Evolution (적응진화를 활용한 cellobiose와 xylose 동시발효 Pichia stipitis의 개발)

  • Kim, Dae-Hwan;Lee, Won-Heong
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.565-573
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    • 2019
  • Production of biofuels and value-added materials from cellulosic biomass requires the development of a microbial strain capable of efficiently fermenting mixed sugars. In this study, the natural xylose fermenting yeast, Pichia stipitis, was evolved to simultaneously ferment cellobiose and xylose. Serial subcultures of wild-type P. stipitis in 20 g/l cellobiose were performed to increase the rate of cellobiose consumption. A total of ten rounds of the serial subculture led to the isolation of an evolved strain fermenting cellobiose significantly faster than the parental strain. The evolved strain displayed enhanced ethanol yield from 0 to 0.4 g ethanol/g cellobiose. The evolved P. stipitis simultaneously fermented cellobiose and xylose in batch fermentation. The genetic information of our evolved P. stipitis would be valuable in the development of a microbial host for the production of biofuels and biomaterials from cellulosic biomass.

Callus Induction from Seeds of Birdsfoot trefoil and Plant Regeneration on BOi2Y Medium (버즈풋 트레포일 종자로부터 캘러스 유도 및 BOi2Y 배지에서 식물체 재분화)

  • Kim, Ki-Yong;Rim, Yong Woo;Choi, Kee Jun;Sung, Byung Ryul
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.4
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    • pp.303-308
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    • 1999
  • The conditions for callus formation and plant regeneration were confirmed in birdsfoot trefoil (Lotus corniculatus L.). Among SH (Schenk and Hildebrandt), MS (Murashige and Skoog) and N6 medium (Chu), SH medium was highest degree of efficiencies respectively in callus formation and plant regeneration. In this study, we determined volume of hormones and other compounds appended in media. For callus formation, only $3mg/{\ell}$ of 2,4-D (2,4-dichlorophenoxy acetic acid) was appended in their media. For plant regeneration, we used BOi2Y medium (Bingham et al.). We obtained birdsfoot trefoil plants from callus by regeneration, about sixty days later transfer calli to regeneration media.

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Isolation and characteristics of yellow-pigment producing mutants of Monascus anka. (Monascus anka로부터 황색소 생성 변이주의 분리 및 특성)

  • 이호재;이형주
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.111-115
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    • 2002
  • To produce yellow pigment selectively, mutants were induced from Monascus anka Nakazawa et Sato IFO 4478 (KCCM 11832 strain), and their characteristics were evaluated. Five kinds of auxotrophic mutants which required amino acids for growth and pigmentation, were isolated through a series of mutagenic treatments. Especially, asparagine auxotroph Y7 produced high ratio of yellow pigment. This mutant showed all the morphological characteristics of Monascuceae but the shape of colony and the diameter of conidia. Mutant Y7 was propagated by sexual reproduction more often than asexual reproduction, which could be effective in production of pigments. Yellow pigment produced extracellularly by the mutant Y7 was more soluble in polar solvents such as ethanol and water than in nonpolar solvents. Its productivity of yellow pigment was 2.2 times higher in the mutant Y7 than in parents. In addition, its yellow pigment showed characteristics of maximum absorption at 373 nm. Moreover, the hue of pigment produced by the mutant Y7 was bright yellow, and it was stable through the subculture over 10 generations.

Studies on Pythium spp. in Korea -(I) Preliminary taxonomic and physiological studies- (한국(韓國)의 Pythium spp. 에 관(關)한 연구(硏究) -(I) 분류(分類) 및 생화학적(生理學的) 기초연구(基礎硏究)-)

  • Lee, Eung-Kwon;Lee, Young-Hee;Yoo, Jae-Dang;Long, Peter G.
    • The Korean Journal of Mycology
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    • v.3 no.2
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    • pp.7-12
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    • 1975
  • Three species of Pythium previously not recorded in Korea were found during 1975. Pythium spinosum Sawada was isolated from diseased cabbage seedlings, P. myriotylum Dreschl. was isolated from kidney bean and P. butleri Subramanium from cucumber, spinach, red bean and radish. Pathogenicity of isolates of P. butleri and P. myriotylum was confirmed in pathogenicity tests but P. spinosum appeared to be non-pathogenic. Several isolates failed to sporulate satisfactorily on cornmeal agar and some produced degenerate oogonia after sub-culturing on this medium. Sunflower seed agar was found to be a suitable alternative medium. The validity of previous records of P. debaryanum Hess in Korea is discussed.

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Physiological properties and transformation of alkaline-tolerant bacteria (알카리내성 세균의 생리적 특성 및 형질전환)

  • 유주현;정용준;정건섭;오두환
    • Microbiology and Biotechnology Letters
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    • v.14 no.3
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    • pp.239-244
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    • 1986
  • To develop the potential use as new host strain for gene cloning, alkaline-tolerant isolates from soil were examined for amylase activity, protease activity, antimicrobial activity and transformability by using plasmid pUB 110. Of these strains, one was selected and identified as Bacillus sp. YA-14. in the enzymatic properties of Bacillus sp. YA-14 the optimal conditions for the reaction of amylase and protease were at pH 0.8 and pH 7.5 respectively. The antimicrobial activity of Bacillus sp. YA-14 was also found. For the transformation, Bacillus sp. YA-14 was cultured to late logarithmic growth phase ai 37$^{\circ}C$ in modified SPI medium (pH 8.0) containing 0.4% MgSO$_4$. The presence of pUB 110 plasmid DNA in transformants was confirmed by electrophoresis and stably maintained in the new host.

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Promotion of in vitro shoot proliferation in rose by addition of liquid medium to culture (액체배지 첨가에 의한 장미 기내 신초 증식 촉진)

  • Lee, Ye Ji;Lee, Jung Lim;Hyung, Nam-In;Kim, Seung Tae;Lee, Eun Kyung;Kwon, O Hyeon;Kim, Won Hee;Lee, Su Young
    • Journal of Plant Biotechnology
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    • v.39 no.4
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    • pp.305-308
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    • 2012
  • To promote the growth and proliferation of in vitro rose (Rosa hybrida L) shoots, a liquid medium was added to shoot culture. Shoots were obtained by culturing internodes of four cultivars, 'Antique Curl', 'Shiny Orange', 'White Zen', and 'Red Zen', and then were proliferated by the subculture two times. An addition with 10~15 mL of liquid medium enhanced the shoot elongation of all four cultivars. However, the effect of liquid medium addition to culture of in vitro shoot for proliferation was dependent on cultivars of rose.

Thiamin Requirements for Vegetative Growth and Fruit Body Formation of Lentinula edodes

  • Shin, Gab-Gyun;Meguro, Sadatoshi;Kawachi, Shinsaku
    • Journal of the Korean Wood Science and Technology
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    • v.28 no.1
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    • pp.48-54
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    • 2000
  • The effects of thiamin on vegetative mycelial growth and fruit body formation of Lentinuia edodes were investigated in basal peptone-glucose liquid medium in relation to the uptake of thiamin. Thiamin was essential for fruit body formation, and the minimum requirements for thiamin were estimated to be approximately 10 ${\mu}g$/L. The vegetative mycelial growth was little influenced by the addition of thiamin in the range of 1.5 ${\mu}g$~1.5 mg/L. While the mycelium was successively transferred to fresh peptone-glucose-agar medium three times, the repression of mycelial growth was not significant. Even in cases using vitamin-free casamino acid or glutamic acid as a nitrogen source instead of peptone, a thiamin deficiency for mycelial growth did not occur as a result of transferring the mycelia to fresh media. Almost all of the thiamin contained in the media accumulated in the mycelia during the first 3 weeks of a 9-week incubation. These results suggest that only trace amounts of thiamin are required for vegetative mycelial growth in Lentinula edodes and that almost all thiamin added to a basal medium will be used for fruit body formation.

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Comparative Effect of Plant Growth Regulators on Callus Induction and Plant Regeneration in Glehnia littoralis Schmidt et Miquiel (갯방풍(Glehnia littoralis Schmidt et Miquiel)에서 캘러스 유도 및 식물체 재분화에 미치는 생장조절제의 효과)

  • 이병국;한미숙;정양균;나의식;윤성중;유남희
    • Korean Journal of Plant Resources
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    • v.17 no.2
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    • pp.153-160
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    • 2004
  • Glehnia littoralis is known as an edible and medicinal plant using green loaves and mature roots of plant. In the present paper, the influence of plant growth regulators on callus induction and plant regeneration was investigated. Callus induction and regeneration occurred from leaf and petiole explants in Glehnia littoralis. Optimal condition of plant growth regulators for callus induction from leaf and petiole explants was MS basal medium supplemented with 2mg/L 2,4-D and 2mg/L BA. The frequency of callus induction was higher in petiole explant than leaf. When the callus was cultured on MS basal medium supplemented with 0∼1 mg/L IAA, 0∼1mg/L NAA and 0∼2mg/L BA for about 65 days, the most effective plant growth regulators on plant regeneration from callus were 1mg/L NAA and 2mg/L BA. The plantlets acclimatized successfully and grown in vermiculite matrix.