• Title/Summary/Keyword: Media Clone

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The Investigation of Cell Culture Conditions to Maintain Chicken Embryonic Stem Cells as Totipotent Cells

  • Du, Lixin;An, Jing
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.8
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    • pp.1102-1107
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    • 2003
  • The ES cell can provide a useful system for studying differentiation and development in vitro and a powerful tool for producing transgenic animalds. To investigate the culture condition of chicken embryonic stem (CES) cells which can retain their multipotentiality or totipotency, three kinds of feeder layer cells, SNL cells, primary mice embryonic fibroblasts (PMEF) cells and primary chicken embryonic fibroblasts (PCEF) cells, were used as the feeder cells in media of DMEM supplemented with leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and stem cell factor (SCF) for co-culture with blastoderm cells from stage X embryos of chicken. The alkaline phosphatase (AKP) test, differentiation experiment in vitro and chimeric chicken production were carried out. The results showed that culture on feeder layer of PMEF yielded high quality CES cell colonies. The typical CES cells clone shape revealed as follows: nested aggregation (clone) with clear edge and round surface as well as close arrangement within the clone. Strong alkaline phosphatase (AKP) reactive cells were observed in the fourth passage cells. On the other hand, the fourth passage CES cells could differentiate into various cells in the absence of feeder layer cells and LIF in vitro. The third and fourth passage cells were injected into the subgerminal cavity of recipient embryos at stage X. Of 269 Hailan embryos injected with CES cells of Shouguang Chickens, 8.2% (22/269) survived to hatching, 5 feather chimeras had been produced. This suggests that an effective culture system established in this study can promote the growth of CES cells and maintain them in the state of undifferentiated and development, which lays a solid foundation for the application of CES cells and may provide an alternative tool for genetic modification of chickens.

Molecular Cloning of Chitinase Genes Family from Serratia marcescens

  • Song, Young-Hwan;Kweon, Oh-Gun
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.103-110
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    • 1993
  • Total genomic DNA library of Serratia marcescens was prepared by inserting Sau3AI partial digesting fragments(above 5 kb) into the dephosphorylated BamHl site of pUC19. In primary screening, two colonies were selected by observing the halo around E. coli transformants grown on the swollen colloidal chitin media. Secondary screening was performed by soaking two colonies with a few drops of 4-methylumbelleliferryl N-acetyl-$\beta$-D-glucocosaminide(4-MuNGlcNAc). As 4-MuNGlcNAc is a specific, fluorogenic substrate for chitinase, the positive clones produce light fluorescence by the exposure under the long wave U.V. light(360 nm). From genomic DNA library derived from pUC19, we have isolated two different chitinase clones, pCH1(11.0Kb) and pCH2(7.5Kb), which show completely different restriction map to each other. The cross-hybridization of pCH1EA and pCH2 have not revealed any hybridization signals to each other.

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Antibiotic Biosynthesis in bldA-like Mutant of Strptomyces coelicolor (Streptomyces coelicolor blAA-like Mutant에서의 항생물질 생합성)

  • Park, Unn-Mee
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.70-77
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    • 1994
  • The author isolated 7 mutant candidates which mapped around cysA (which was 10 o'clock). They were divided into two groups. One of them was located counterclockwise to cysA, and the other was clockwise to cysA. Since bldA was mapped counterclockwise to cysA, the candidate which mapped counterclockwise to cysA was transduced with phage containing wild type bldA gene clone. The candidates might be the alleles of bldA, because they were complemented by bldA clone. However some of such mutants sporulated very well and developed as much pigment as wild type on rich media plate. Their phenotype was not like bld mutant at all on such conditions. There were real antibiotics gene expressions, since transcriptional reporter gene xylE had shown high activities. Majority of the bldA like mutants showed act gene expressions when they were transformed with high copy number plasmid containing actII-ORF4.

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Transfer of Insecticidal Toxin Gene in Plants: 2. Subcloning of B. thuringiensis Insecticidal Protein Gene and Rapid Plantlet Regeneration from Nicotiana tabacum Protoplast and Callus (식물세포에 살충독소유전자의 전이연구: 2. B. thuringiensis 살충독소유전자의 Subcloning과 Nicotiana tabacum의 원형질체와 칼루스로부터 신속재생연구)

  • 이형환;조상현황성희김수영
    • KSBB Journal
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    • v.6 no.3
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    • pp.289-297
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    • 1991
  • The insecticidal protein gene in the pKL-20-1 clone derived from Bacillus thuringiensis serovar. kurstaki plasmid was subcloned in the plant shuttle vector, pGA643. The 7.3 kb fragment was cloned in the BglII and Hpal sites of pGA643 vector and expressed in E. coli S17-1, which produced insecticidal proteins killing Bombyx mori larvae. The clone was named pHL-20. The protoplast formation, calli induction and plantlet regeneration of Nicotiana tabacum was carried out. A tremendous number of mesophyll protoplasts of N. tabacum were formed, up to 7$\times$105 protoplast per ml, for 20 hours in darkness in the enzyme solution of 0.5% cellulase and 0.1% macerosin, pH 5.8. The viabilities of the protoplasts were maintained above 80% for 6 days in the media containing 2mg/1 of NAA and 1mg/1 of kinetin. Calli were induced from the protoplasts and leaves of the N. tabacum on MS medium containing 0.5mg/1 BAP. Under the culture conditions the protoplasts underwent repeated cell division into calli. Plantlets were regenerated from callus cultures derived from protoplast and leaves. Shoots were induced in a medium containing 1mg/1 of BAP.

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Promotion of Rootability of the Cuttings of Dendropanax morbifera Lev (황칠나무의 揷木發根力 增進)

  • 김세현
    • Korean Journal of Plant Resources
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    • v.11 no.2
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    • pp.157-162
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    • 1998
  • This study was conducted to examine the effects of media, crown position, clone ,a nd plant growth substances upon the rootability of cuttings for the establishment of the method of mass production by cutting of Dendropanax morbifera Lev. the artifical soil mixtureof vermiculite, perlite and peatmoss (1 : 1 : 1 v/v/v) with IBA 100mg /l treatment was most effective on the rooting rate (85%) was obtained with semihardwood cuttings, which were collected early-August after the first growing seasons. The effect of crown position on rooting of cuttings were obsreved. Average percentage rooting of cuttings on crown position showed 78% , 72% and 65% in middle , upper and lower part with IBA 100mg/l treatment , respectively. Among the four treatment levels of IBA, the highest rooting rate was observed in 100mg/l IBA treatment.

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Establishment of 8-Azaguanine Resistant Human Plasmacytoma Cell Line (8-Azaguanine 내성 인형 형질세포종 세포주의 확립)

  • Cha, Chang-Yong;Hwang, Eung-Soo;Kook, Yoon-Ho;Lim, Dong-Kyun;Cho, Han-Ik;Park, Myung-Hee;Kim, Noe-Kyung;Chang, Woo-Hyun;Lee, Mun-Ho
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.3
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    • pp.399-406
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    • 1986
  • This study was performed to establish human plasmacytoma cell line as the partner cells for producing human hybridoma. Bone marrow cells from a multiple myeloma patient from Seoul National University Hospital, Korea were cultured and established as the cell line, named as HMC-BM4. HMC-BM4 cells were cultivated in RPMI 1640 media containing 8-azaguanine(8-AG; gradually increasing concentration from $1\;{\mu}g/ml$ to $20\;{\mu}g/ml$). 8-AG resistant cells were collected and cloned by limiting dilution. Each clone was divided and tested to die in hypoxanthine, aminopterine and thymidine (HAT) selection media. Finally one clone was selected and named as HMC-AR, which was sensitive to HAT selection media. HMC-AR cells showed typical morphology of plamacytoma in Wright staining. No cell formed the rosette with sheep erythrocytes. Surface membrane $\mu$ heavy chain was detected in 20% of HMC-AR cells and cytoplasmic $\mu$ heavy chain in 90% of them by direct immunofluorescent staining. Ia-like antigen was found in 90% of HMC-AR cells by indirect immunofluorescent staining using anti-Ia-like antigen monoclonal antibody, 1BD9-2. And about $1.0\;{\mu}g/ml$ of human $\mu$ heavy chain was detected in the 3-day culture supernatant of HMC-AR cells. 88% of cells contained 46 chromosomes. Mycoplasma was not detected in HMC-AR cells by Hoechst 33258 staining. This cell line would be used for making hybridomas secreting human monoclonal antibody.

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Cloning of a Chitinase Gene of Xanthomonas sp. Isolated from Soil and its Expression in E. coli. (토양에서 분리된 Xanthomonas sp.의 Chitinase 유전자 cloning과 E.coli에서의 발현)

  • Kim, Ho-Sang;Seong, Ki-Young;Eun, Moo-Young;Hwang, Cher-Won
    • Applied Biological Chemistry
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    • v.41 no.2
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    • pp.125-129
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    • 1998
  • Xanthomonas sp. isolated from soil exhibited cell wall lytic activity of Candida albicans and secreted chitinase in chitin media. Especially, the chitinase activity was induced by chitin and reached a maximum level at 3 days culture in chitin media. We constructed genomic library of Xanthomonas sp. using cosmid vector in E. coli. Oligonucleotide probe was synthesized from the consensus sequence corresponding to chitinase active site, which was derived from the comparison of amino acid sequences of bacterial chitinase genes. Using this oligonucleotide probe, we screened the genomic library. By restriction enzyme mapping of the positive clones, we identified 4 independent clones which may contain the chitinase gene. One of the clones, named pXCH1 (1.2 kb insert), was further analyzed. Northern blot analysis indicated that is transcripts, 1 kb and 0.8 kb, were induced by chitin. When the cloned gene was induced by IPTG in E.coli cell, chitinase activity which was secreted onto culture media was not observed. However, when the cell was disrupted by using sonicator and then centrifuged, the supernatant exhibited chitinase activity. SDS-PAGE of the supernatant indicated that about 35 kDa protein was induced by IPTG. From these results, it was concluded that the cloned DNA was one of the chitinase genes of Xanthomonas sp.

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Mapping of Synchronized Contents with Music to 3 Dimensional Spatial Cloned Human-Object (3차원 공간으로 복제된 인간 형상 오브제에 음악과 동기화한 콘텐츠의 매핑)

  • Bae, Moon-Kyoung;Lyu, Jae-Ha;Kim, Sangwook
    • The Journal of the Korea Contents Association
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    • v.14 no.11
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    • pp.620-627
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    • 2014
  • In the contemporary age, social environment has changed from analog to digital. As a media is developed, the public is more familiar to digital than analog. In this society, media art rises as new art. One of the most useful methods in the media art is projection mapping. In this paper, I propose a realization process of artwork 'Cloned Me' that is realized by projection mapping method. Artwork is formed by projecting image contents that are visualized a music on a steel structure. It is different from media facade that the object projected contents has a story composition. It is a strength that the object and contents are connected by story and composition. Also it can be modified each other easily for harmony because the structure is assembly-type. A face that is projected by beam projection is consist of unique structure not fixed rectangular frame. Artwork 'Cloned Me' has significance that it reflects the present age to develop digital cloning.

Rutin production in hairy root cultures of buckwheat(Fagopyrum esculentum Moench)

  • Park, Sang-Un;Kim, Yeon-Bok;Park, Cheol-Ho
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.23-23
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    • 2002
  • We established hairy root cultures of F. esculentum transformed with A. rhizogenes for in vitro rutin production. Additionally, we describe the effects of different media and plant growth regulators on growth and rutin biosynthesis in buckwheat hairy root cultures. Excised leaves of P. tinctorium from 10-day-old seedlings were used as the explant material for co-cultivation with A. rhizogenes 15834. The hairy culture of Fagopyrum esculentum Moench. was established by infecting leaf explants with Agrobacterium rhizogenes 15834. About four to five weeks after co-cultivation with A. rhizogenes, 10 hairy roots were excised from the necrotic explant tissues. After repeated transfer to fresh medium for three months, ten clones were transferred to MS liquid culture medium. The growth and rutin production of each clone differently response to the MS liquid medium. Among these clones, H8, which had exhibited good growth rate and one of the highest rutin productivity, was selected for the following experimment.(중략)

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Model Inversion Attack: Analysis under Gray-box Scenario on Deep Learning based Face Recognition System

  • Khosravy, Mahdi;Nakamura, Kazuaki;Hirose, Yuki;Nitta, Naoko;Babaguchi, Noboru
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.15 no.3
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    • pp.1100-1118
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    • 2021
  • In a wide range of ML applications, the training data contains privacy-sensitive information that should be kept secure. Training the ML systems by privacy-sensitive data makes the ML model inherent to the data. As the structure of the model has been fine-tuned by training data, the model can be abused for accessing the data by the estimation in a reverse process called model inversion attack (MIA). Although, MIA has been applied to shallow neural network models of recognizers in literature and its threat in privacy violation has been approved, in the case of a deep learning (DL) model, its efficiency was under question. It was due to the complexity of a DL model structure, big number of DL model parameters, the huge size of training data, big number of registered users to a DL model and thereof big number of class labels. This research work first analyses the possibility of MIA on a deep learning model of a recognition system, namely a face recognizer. Second, despite the conventional MIA under the white box scenario of having partial access to the users' non-sensitive information in addition to the model structure, the MIA is implemented on a deep face recognition system by just having the model structure and parameters but not any user information. In this aspect, it is under a semi-white box scenario or in other words a gray-box scenario. The experimental results in targeting five registered users of a CNN-based face recognition system approve the possibility of regeneration of users' face images even for a deep model by MIA under a gray box scenario. Although, for some images the evaluation recognition score is low and the generated images are not easily recognizable, but for some other images the score is high and facial features of the targeted identities are observable. The objective and subjective evaluations demonstrate that privacy cyber-attack by MIA on a deep recognition system not only is feasible but also is a serious threat with increasing alert state in the future as there is considerable potential for integration more advanced ML techniques to MIA.