• Title/Summary/Keyword: 계대

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In vitro propagation of oil palm (Elaeis guineensis Jacq.) clones through somatic embryogenesis and analysis of somaclonal variation by RAPD (체세포배발생을 통한 오일팜나무(Elaeis guineensis Jacq.) 클론의 기내증식 및 RAPD를 이용한 체세포변이의 검정)

  • Ahn, In-Suk;Park, Hye-Rim;Son, Sung-Ho
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.196-204
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    • 2012
  • This study was carried out to develop reliable systems for somatic embryogenesis in oil palm tree (Elaeis guineensis Jacq.), and to verify the somaclonal variants by RAPD analysis. Embryogenic callus was induced successfully on modified half-strength MS medium containing $NaH_2PO_4{\cdot}2H_2O$ and casein. Embryogenic callus was further developed to somatic embryo mass (SEM), which is very hard and bonded tightly each other. Plantlets were proliferated when SEM was cultured on modified MS medium containing half strength $NH_4NO_3$, casein and L-ascorbic acid. Plantlets were transplanted into pots containing artificial soils. When RAPD analysis was conducted using randomly selected 95 in vitro plantlets and 19 random primers, somaclonal variation was detected using BNR35 primer. There was missing band around 1 kb in #22, #28, #35, and #77 plantlets. In addition, bands obtained from #28, #35, and #77 was much stronger than other normal bands. The blast results at NCBI revealed that somaclonal variation observed in this study was related to chloroplast genome of oil palm. The results also revealed that oil palm reproduction system through somatic embryogenesis is quite reliable and early detection of somaclonal variants seem to be possible at in vitro stage by RAPD analysis.

Isolation of porcine reproductive and respiratory syndrome virus(PRRSV) in Korea (돼지생식기 및 호흡기증후군(Porcine Reproductive and Respiratory Syndrome ; PRRSV) 바이러스의 국내분리주 작성에 관한 연구)

  • Kweon, Chang-hae;Kwon, Byung-joon;Lee, Han-jung;Cho, Jae-jin;Hwang, Eui-kyung;Shin, Jin-ho;Yoon, Yong-dhuk;Kang, Yung-bai;An, Soo-hwan;Kim, Yong-hee;Huh, Won;Jun, Moo-hyung;Wensvoort, G.
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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    • pp.77-83
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    • 1994
  • Three viral strains, causing CPE in porcine alveolar macrophage cell, were isolated from aborted fetus, serum from young pig showing blue-ear sign and lung of suspected pig, respectively. The differential diagnostic results showed no characteristics of Aujeszky's disease virus(ADV), hog cholera virus (HCV), Japanese encephalitis virus(JEV), porcine parvovirus(PPV) and encephalomyocarditis virus (EMCV). However, positive reactions were demonstrated by IFA using monospecific porcine antibodies against Lelystad virus. When the paired sera of experimentally inoculated swine with one of isolate, KPRRSV-l were tested by IPMA, the result indicated that the isolate was related to United States isolate than European LV.

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Characterization of cultures isolated from fruiting body tissue in Armillaria gallica (천마버섯(Armillaria gallica) 자실체 조직배양체의 특성)

  • Yoo, Young-Bok;Oh, Jin A;Oh, Youn-Lee;Moon, Jiwon;Shin, Pyung-Gyun;Jang, Kab-Yeul;Kong, Won-Sik
    • Journal of Mushroom
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    • v.11 no.2
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    • pp.63-68
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    • 2013
  • The fruiting body of honey mushroom, Armillaria gallica, was collected from Gastrodia elata cultivated fields. Pure cultures were isolated from fruiting body tissue of the mushrooms, and cultured on MCM (mushroom complete medium) or PDA (potato dextrose agar) medium. Then, 12 different types of mycelial growth characteristics such as growth rate, colony morphology and rhizomorph formation were obtained. The vitality of the mycelial growth and rhizomorph formation of the fruiting body culture isolates were better on MCM than PDA, suggesting that the optimal culture medium for A. gallica mycelia was MCM. To observe the feature of colony morphology, the subculture of isolates were incubated on MCM. Consequently, we could find the segregated or differentiated colony morphology from isolate type 11 that was similar morphology to isolate type 12. For phylogenetic analysis of the 12 isolates, RAPD (Random Amplified Polymorphic DNA) were performed. The isolate type 12 was not only shown different band patterns of RAPD variation in other 11 isolates, but also commercial strain known as Chunmagyun No. 1. Among the tissue culture isolates of fruiting, strains with better mycelial growth characteristics than Chunmagyun No. 1 were selected. We expect that the new strain can be substituted to commercial strain Chunmagyun No. 1.

Production and Characterization of a Transgenic Mouse Model of Human Liver Cancer (형질 전환 기법을 이용한 인체 간암의 마우스 모델 제작 및 특성 규명)

  • Li, Zhong-Shu;Lee, Jung-Woong;Hyun, Byung-Hwa;Lee, Chul-Ho;Jeong, Kyu-Shick;Fang, Nan-Zhu;Yeom, Young-Il
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.145-152
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    • 2007
  • Transgenic mice were generated by microinjecting a plasmid DNA containing the SV40 (simian virus 40) large T antigen (Tag) gene fused with mouse albumin promoter/enhancer sequences into fertilized one-cell mouse embryos. Among eleven founder transgenic animals, four developed hepatocellular carcinoma, two showed kidney cancer and one developed skin and brain tumors. Three stable transgenic lines, #1-2, #1-6 and #1-11 were established. Members of the lines #1-6 and #1-11 reproducibly developed liver tumors by 8 to 10 weeks of age but did not exhibit any phenotypic changes in other tissues. Histological changes loading to liver tumor formation occurred with predictable kinetics and could be classified into three distinct stages; (a) newborn to 3 weeks of age, characterized by hyperplastic hepatocytes with reduced amounts of cytoplasm without any nuclear alterations, (b) between 4 to 8 weeks of age, characterized by diffuse liver cell dysplasia without observable tumor nodules, and (c) 9 weeks of age and thereafter, characterized by hepatocellular carcinomas in the background of extensive liver dysplasia. Metastasis to the lung from a liver carcinoma was observed in #1-11 founder animal. This transgenic mouse system displays similarities with human liver cancers in a number of aspects and provides a useful model for the study of molecular events involved in hepatocarcinogenesis.

Anti-wrinkle Activity of Low Molecular Weight Peptides Derived from the Collagen Isolated from Asterias amurensis (불가사리(Asterias amurensis) 콜라겐 유래 저분자 펩타이드의 피부주름 억제활성)

  • Kwon, Min-Chul;Kim, Cheol-Hee;Kim, Hyo-Sung;Syed, Abdul Qadir;Hwang, Bo-Yong;Lee, Hyeon-Yong
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.625-629
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    • 2007
  • This study was carried out to investigate the anti-wrinkle effects of peptides derived from collagens isolated from Asterias amurensis, which was collected in the East Sea. The molecular weights of the peptides were between 10-50 kDa, as determined through sephadek G-75 gel. The cytotoxicities against CCD-986sk cells and HEL-299 cells were measured using the MTT assay. The cytotoxicity of all the fractions(F1: Fraction No. 4-13, 116 kDa; F2: Fraction No. 25-30, 100 kDa; F3: Fraction No. 45-55, 58 kDa; F4: Fraction No. 59-63, 43 kDa; F5: Fraction No. 79-90, 24 kDa) was less than 25%, by the addition of 1.0 mg/mL. These peptides did not show any adverse effects on human skin cells. In the presence of F1 at 1.0 mg/mL, matrix metalloproteinase-1 (MMP-1) expression of UVA-induced human normal fibroblasts was reduced to 34.8%. Overall, the results seem to suggest that peptides of approximately 20 kDa have superior anti-wrinkle effects.

Effects of EGF, $\beta-ME$, Glucose on the In Vitro Maturation and Development of Porcine NT Embryos (EGF, $\beta-ME$와 Glucose가 돼지 난자의 체외성숙에 미치는 영향과 핵 이식 배의 발생에 대한 영향에 관한 연구)

  • Quan J. H.;Kim S. K.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.137-145
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    • 2005
  • 본 연구는 NCSU-23과 PZM-3 배양액에 EGF, $\beta-ME$와 glucose의 첨가가 돼지 난자의 체외성숙에 미치는 영향과, 배양조건을 다르게 하여 계대배양한 섬유아세포를 이용한 핵이식 배를 다른 배양액과 산소조건에서 배양하였을 때 체외발생율에 미치는 영향을 조사하였다. 난자를 20ng/ml EGF를 첨가 또는 첨가하지 않은 NCSU-23 및 PZM-3 배양액에서 44시간 배양하였을 때 난자의 체외성숙율은 각각 $85.7\pm3.1\%,\;75.2\pm2.8\%,\;87.1\pm2.4\%$$78.2\pm2.6\%$였으며 EGF를 첨가한 배양액에서 배양한 난자의 체외성숙율은 EGF를 첨가하지 않은 배양액에서 배양했을 때의 난자보다 높은 체외성숙율을 나타냈다(p<0.05). 난자를 $25{\mu}M\;\beta-ME$를 첨가 또는 첨가하지 않은 NCSU-23 및 PZM-3 배양액에서 44시간 배양하였을 때 난자의 체외성숙율은 각각 $79.5\pm2.6\%,\;74.7\pm2.5\%,\;80.2\pm2.3\%,\;78.6\pm2.7\%$였고 $\beta-ME$를 첨 가한 PZM-3 배양액에서 배양한 난자의 체외성숙율이 가장 높게 나타났다. 난자를 1.5mM glucose를 첨가 또는 첨가하지 않은 NCSU-23 및 PZM-3 배양액에서 44시간 배양하였을 때 난자의 체외성숙율은 각각 $79.2\pm2.3\%,\;75.0\pm2.6\%,\;85.5\pm2.5\%$$78.9\pm2.7\%$였고, glucose를 첨가한 PZM-3 배양액에서 배양한 난자의 체외성숙율은 glucose를 첨가하지 않은 PZM-3 배양액에서 배양한 난자보다 높은 체외성숙율을 나타냈다(p<0.05). 핵이식 배를 20ng/m1 EGF, $25{\mu}M\;\beta-ME$ 및 1.5mM glucose를 첨가한 NCSU-23 및 PZM-3 배양액에서 48시간, 144시간 배양하였을 때 2세포기 및 배반포로의 체외발생율은 각각 $56.4\pm2.7\%,\;54.3\pm2.9\%,\;70.5\pm2.1\%,\; 69.6\pm1.5\%$$12.0\pm1.3\%,\;9.6\pml.7\%,\;10.9\pm2.1\%,\;11.9\pm1.8\%$였다.

Effect of Thidiazuron on Callus and Multiple Shoot Formation in Shoot-tip Culture of Hibiscus syriacus L. 'Honghwarang' (Thidiazuron이 무궁화 '홍화랑' 품종의 정단배양으로부터 Callus형성과 Multiple Shoot형성에 미치는 효과)

  • Kim, Eun Kyoung;Yoo, Yong Kweon;Kim, Ki Sun
    • Horticultural Science & Technology
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    • v.16 no.4
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    • pp.520-524
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    • 1998
  • This study was carried out to investigate the effect of thidiazuron(TDZ) on callus and shoot primordia formation, to determine the most optimum multiple shoot induction medium, and to obtain the plantlets on solid medium via shoot organogenesis. TDZ 0.01 mg/L in MS medium was most effective on callus formation, and BA 0.1 mg/L was most effective on shoot growth, while TDZ 0.01 mg/L was most effective on callus formation. TDZ 0.001 mg/L was most effective in shoot primordia formation. Shoot tips were cultured with TDZ 0.01 mg/L for 8 weeks and induced callus was transferred to regeneration medium containing TDZ 0.001 mg/L. After 4 weeks induced shoot primordia were resubcultured at growth regulator-free medium for 4 weeks. The induced multiple shoots rooted more efficiently at NAA 1.0, 5.0 mg/L, or IBA 5.0 mg/L.

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Growth Promotion of Pavlova viridis by Bacteria Isolated from the Microalga (파블로바 비리디스로부터 분리한 세균에 의한 미세조류의 생장 촉진)

  • Ahamed, Sarker Anowarul Kabir;Kim, Jin-Joo;Choi, Tae-O;Choi, Tae-Jin
    • Journal of Life Science
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    • v.25 no.5
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    • pp.568-576
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    • 2015
  • The marine microalga Pavlova viridis can grow fast and has the ability to accumulate essential nutrients for culturing marine animals, such as EPA and DHA, and it has been used as food for raring larval fish and prawn. The symbiotic relationship between the flagellate microalga Pavlova viridis and its associated bacteria was investigated. An axenic culture of P. viridis was obtained by repeated treatment of the microalga with an antibiotic cocktail. The axenic status was confirmed after sub-culturing three times in a sterile f/2 medium without an antibiotic. The axenic alga was then co-inoculated with five bacteria, arbitrarily designated as I1–I5, isolated from the alga to test the growth promotion of the algae. All bacterial strains promoted the growth of P. viridis, and bacterial isolate I3 was the most effective among the five bacteria tested. The cell number of P. viridis in the co-culture with I3 was significantly higher than that of the control culture. A sequence analysis of the 16S rRNA gene isolated from I3 revealed a 97% nucleotide sequence similarity to that of Citrobacter sp. The growth of strain I3 was also significantly enhanced by co-culturing with P. viridis, indicating a symbiotic relationship between the microalga and its associated bacterium. The association between the microalga and bacterium was confirmed by scanning electron microscopy.

BACTERIOLOGICAL STUDIES ON MARKET SEA FOODS 1. Sanitary Indicative Bacteria in Sundried Sea Foods. (시판 수산식품에 대한 세균학적 연구 1. 건제품의 위생지표 세균에 관하여)

  • CHANG Dong-Suck;CHOE Wi-Kyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.6 no.3_4
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    • pp.87-91
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    • 1973
  • This study was conducted to evaluate the sanitary quality of sun dried sea foods being distributed in the markets located in Busan city. Twenty one kinds of sun dried sea foods, 9 kinds of fish and shellfish, 5 kinds of mollusca and 7 kinds of seaweed were examined during the Period from September to December in 1970. For the evaluation of sanitary contents of sanitary indicative bacteria such as coliform group, fecal coliform, fecal streptococci and enterococci, and plate counts were determined. The results obtained were as follows: 1. Through out all the samples examined, the numbers of fecal streptococci and enterocccci were much greater than those of coliform group and fecal coliform. 2. Generally, the contents of enterococci exceeded MPN of 1,000 per 100 grams through all samples examined, while fecal coliform MPN'S were less than 18. 3. The median value of coliform group MPN was 78 and those of fecal streptococci MPN was 3,300 per 100 grams, The median value of the plate counts was $9.9\times10^4$ per gram, 4. Bacterial densities of the foods which had many chances to come into contact with hands were much higher than those of other foods which had not. 5. The bacterial contamination rate of seaweed was much lower than those of fish and shellfish. 6. No correlation between the numbers of sanitary indicative bacteria and plate counts was observed.

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Profiling of genes in healthy hGF, aging hGF, healthy hPDLF and inflammatory hPDLF by DNA microarray (DNA microarray법을 이용하여 건강한 치은섬유모세포, 복제노화된 치은섬유모세포, 건강한 치주인대섬유모세포와 염증성치주인대섬유모세포에서 유전자 발현)

  • Yun, Sang-Jun;Kim, Byung-Ock;Yun, Jeong-Hun;Kang, Dong-Wan;Jang, Hyun-Seon
    • Journal of Periodontal and Implant Science
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    • v.36 no.3
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    • pp.767-782
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    • 2006
  • 이 연구의 목적은 DNA microarray 분석법을 이용하여 건강한 사람치주인대섬유모세포, 건강한 사람치은섬유모세포, 복제노화된 사람치은섬유모세포, 염증성 사람치주인대 섬유모 세포의 유전자 발현 형태를 상호비교하고자 하였다. 환자의 동의하에 충치, 치주염이 없이 교정발치된 치아의 치주인대세포를 배양하여 건강한 치주인대섬유모세포로, 만성치주염으로 발거된 치아에서 채취하여 배양한 세포를 염증성 치주인대섬유모세포로 선정하였다. 구강에서 채취한 치은결체조직에서 배양한 사람치은섬유모세포를 일차 배양한 후 계대배양을 통해 복제 노화를 유도하였다. $-198^{\circ}C$의 액화질소에 저장되어 있던 2, 4, 8, 15, 16세대 세포를 실험에 이용하였다. 위의 모든 세포들은 60 mm 배양접시에서 세포들이 80-90%의 밀생이 될 때까지 5% $CO_2$, $37^{\circ}C$, 100% 습도의 배양기에서 2일 간격으로 10% FBS가 함유된 DMEM 세포 배양액을 교체하면서 세포를 배양하였다. Trizol Reagent (Invitrogen, USA)를 이용하여 제조회사의 지시에 따라 total RNA를 추출하였다. 18S RNA와 28S RNA를 확인한 후 DNA microarray 분석을 실시하였다. 4배수 이상의 변화양상을 비교시 상호 유전자 발현의 차이를 나타내었다. 건강한 사람치은섬유모세포(2세대)와 노화된 치은섬유모세포에서 가장 높은 발현변화를 나타낸 반면 DMC1 dosage suppressor of mck1 homolog, meiosis-specific homologous recombination,은 건강한 치은섬유모세포에서 가장 높게 나타났다. 염증성 치은인대섬유모세포와 건강한 치주인대 섬유모세포를 비교시, Regucalcin은 염증성 치주인대섬유모세포에서 가장 높게 나타났고, Vascular cell adhesion molecule 1도 두 번째로 높게 나타났다. 건강한 치주인대섬유모 세포와 건강한 치은섬유모세포를 비교시, IL-11과 periostin이 치주인대섬유모세포에서 높은 발현을 나타낸 반면, Prostaglandin D2 synthase 21kDa과 Thioredoxin interacting protein은 치은섬유모세포에서 높은 발현을 나타내었다. 염증성 치주인대섬유모세포와 노화된 치은섬유모세포(15세대 이상)를 비교시 149개의 유전자가 유사한 발현 수준을 나타내었다. 이 연구는 노화, 염증, 세포 형태에 따라서 유전자 수준에서 가장 높거나 높은 수준 변화를 보이는 유전자가 다를 수 있음을 나타낸다. 향후, 치주염 환자들에서, 노염, 염증, 세포 특이성에 관한 유전자 표시지를 이용하여 진단하거나 치료에 응용하기 위해서는 더 많은 연구가 필요하리라 사료된다.