• 제목/요약/키워드: Media Clone

검색결과 40건 처리시간 0.023초

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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    • 제16권8호
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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
    • 한국어병학회지
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    • 제6권2호
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    • pp.103-110
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    • 1993
  • Sau3AI으로 부분절단한 Serratia marcescens genomic DNA(5Kb 이상)을 pUC19의 BamHI site에 삽입하여 total genomic library를 준비하였다. Swollen colloidal chitin media에서 halo를 형성하는 2개의 E.coli 형질전환주를 선별하였다. 이들 colony가 chitinase 유전자를 갖음을 재확인하기 위하여 4-methylumbelliferyl N-acetyl-$\beta$-D-glucosaminide(4-MuFGlcNAc)를 이용하였다. 4-MuFGlcNAc는 chitinase에 대한 기질특이성을 나타내며 형광을 나타내는 기질로서 positive clone들은 360nm의 자외선을 조사하였을 경우 밝은 형광을 나타낸다. pUC19으로 부터 유래된 2 종류의 다른 chitinase clone, pCH1(11.0Kb) 및 pCH2(7.5Kb)를 genomic DNA library로 부터 분리하였으며, 이들의 제한효소지도를 작성한 결과 서로 다른 제한효소지도를 나타내었다. pCH1EA 및 pCH2로 부터 각각의 EcoRI-Xbal fragment를 subcloning함으로써 두개의 다른 chitinase 유전자의 위치를 결정하였다. pCH1EA 및 pCH2를 cross hybridization 한 결과 hybridization signal을 나타내지 않아 서로 유사성이 없는 것으로 사료된다.

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

  • 박은미
    • 미생물학회지
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    • 제32권1호
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    • pp.70-77
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    • 1994
  • Streptomyces coelicolor를 화학적 방법으로 돌연변이시킨 결과, 유전적 교잡방법에 의해 bldA와 유사한 위치인 10시 방향으로 위치가 결정되는 변이주들을 찾아내었다. 이들의 자세한 위치를 점검한 결과 cysA를 중심으로 반시계 방향으로 떨어져 있는 group고가 시계방향으로 떨어져 있는 두가지로 나뉘었다. 반시계 방향으로 떨어져 있는 변이주들을 bldA와 유사한 변이주라고 생각되어 이를 확인하기 위해 정상적인 bldA 유전자가 cloning 되어 있는 phage를 이용하여 기능적인 complementation 여부를 확인하여 보았다. Complementation이 되는 것으로 미루어 보아 이 변이주들이 bldA의 allele일 가능성이 매우 높으나 이들 중 몇몇 변이주들은 기존의 bldA와 무척 다른 양상을 보였다. 즉, 고영양 배지인 $R_2$YE배지에서 왕성하게 spore를 생산하며 wild type이 생산하는 이상의 색소 생산을 보인 점이다. 이때까지 발견된 bldA 변이주들은 배지 조성에 따라 약간의 aerial mucelium을 형성하는 것이 보고되었으나 이렇게 조건에 따라서는 완전히 bld phenotype을 잃는 변이주는 보고되지 않았다. 이 색소 생산이 실지로 항생물질 유전자의 발현에 의한 것이라는 것을 xylE fusion을 이용했을 때 유전자의 전사가 일어나는 것이 확인됨으로써 증명되었다. 또 이들 bldA 유사 변이주들은 actII-ORF4가 높음 copy수로 들어있는 pasmid에 의해 act 유전자를 발현하는 것도 관찰되었다.

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식물세포에 살충독소유전자의 전이연구: 2. B. thuringiensis 살충독소유전자의 Subcloning과 Nicotiana tabacum의 원형질체와 칼루스로부터 신속재생연구 (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)

  • 이형환;조상현황성희김수영
    • KSBB Journal
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    • 제6권3호
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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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황칠나무의 揷木發根力 增進 (Promotion of Rootability of the Cuttings of Dendropanax morbifera Lev)

  • 김세현
    • 한국자원식물학회지
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    • 제11권2호
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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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8-Azaguanine 내성 인형 형질세포종 세포주의 확립 (Establishment of 8-Azaguanine Resistant Human Plasmacytoma Cell Line)

  • 차창룡;황응수;국윤호;임동균;조한익;박명희;김노경;장우현;이문호
    • 대한미생물학회지
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    • 제21권3호
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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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토양에서 분리된 Xanthomonas sp.의 Chitinase 유전자 cloning과 E.coli에서의 발현 (Cloning of a Chitinase Gene of Xanthomonas sp. Isolated from Soil and its Expression in E. coli.)

  • 김호상;성기영;은무영;황철원
    • Applied Biological Chemistry
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    • 제41권2호
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    • pp.125-129
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    • 1998
  • 한국 토양에서 분리된 Xanthomonas sp.는 Candida albicans에 대한 용균성을 나타내며 분비효소로서 chitinase를 분비하는 것으로 사료되었다. 특히 chitinase활성은 chitin배지에서 배양했을 때 3일 배양에서 최대치를 나타내었다. 이러한 특성이 있는 Xanthomonas의 chitinase 유전자를 cloning하기 위하여 cosmid vector를 이용한 genomic library를 작성하였으며, 다른 박테리아 chitinase 유전자와 homology를 가진 지역의 DNA sequence를 oligonucleotide로 합성하여 probe로 사용한 결과 4개의 독립된 positive clone을 cloning 하였다. 이중 pXCHl(1.2 kb insert) 이라고 명명한 clone에 대해 해석한 결과 이 크론의 전사산물은 chitin 배지에서만 유도됨을 확인하였으며 대장균 발현 vector를 이용한 이 유전자의 대장균에서의 발현에 대한 실험의 결과 약 35 kDa의 단백질을 생산하는 것으로 확인하였다. 또한 이 산물의 chitinase활성을 측정한 결과 유전자가 포함되지 않은 산물에 비해 약 10배의 활성을 나타내어 이 유전자를 Xanthomonas sp.의 chitinase유전자임을 증명하였다.

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

  • 배문경;류재하;김상욱
    • 한국콘텐츠학회논문지
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    • 제14권11호
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    • pp.620-627
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    • 2014
  • 현시대의 사회적 환경은 디지털화되고 있으며, 미디어의 발전으로 대중들은 아날로그보다 디지털에 익숙해지고 있다. 그러한 사회 구조 속에서 대두되고 있는 미디어아트는 다양한 작업방식 중에서 프로젝션 매핑방식이 많이 활용되고 있다. 본 연구에서는 'Cloned Me' 작품을 바탕으로 프로젝션 매핑 작품 구현 과정에 대해 논하고자 한다. 음악을 가시화한 영상 콘텐츠를 제작하고 철 구조물 위에 프로젝션 매핑으로 영상 콘텐츠를 투사하는 방식으로 작품이 구현된다. 스토리를 담은 오브제를 제작하고 그 위에 프로젝션 매핑 방식으로 영상 콘텐츠를 투사함으로써 기존의 미디어 파사드와 차별성을 가진다. 오브제와 콘텐츠 사이를 연결하는 스토리와 구성이 존재하고 상호보완적 수정이 용이하다는 것이 장점이다. 프로젝션이 투사되는 면이 사각형의 고정된 프레임에서 벗어나 독특한 구조를 이룬다. 복제가 가능한 디지털 기술이 발전한 현시대의상을 반영해주는 작품이라는 점에서 의의를 지닌다.

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

  • Park, Sang-Un;Kim, Yeon-Bok;Park, Cheol-Ho
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2002년도 제9차 국제심포지움 및 추계정기학술발표회
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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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    • 제15권3호
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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.