• 제목/요약/키워드: KB-C2

검색결과 471건 처리시간 0.026초

체세포에 있어서 Knock-in 벡터 상동영역 구조에 따른 Knock-in 효율 (Knock-in Efficiency Depending on Homologous Arm Structure of the Knock-in Vector in the Bovine Fibroblasts)

  • 김세은;박다솜;구덕본;강만종
    • Reproductive and Developmental Biology
    • /
    • 제41권1호
    • /
    • pp.7-16
    • /
    • 2017
  • The knock-in efficiency in the fibroblast is very important to produce transgenic domestic animal using nuclear transfer. In this research, we constructed three kinds of different knock-in vectors to study the efficiency of knock-in depending on structure of knock-in vector with different size of homologous arm on the ${\beta}-casein$ gene locus in the somatic cells; DT-A_cEndo Knock-in vector, DT-A_tEndo Knock-in vector I, and DT-A_tEndo Knock-in vector II. The knock-in vector consists of 4.8 kb or 1.06 kb of 5' arm region and 1.8 kb or 0.64 kb of 3' arm region, and neomycin resistance gene(neor) as a positive selection marker gene. The cEndo Knock-in vector had 4.8 kb and 1.8 kb homologous arm. The tEndo Knock-in vector I had 1.06 kb and 0.64 kb homologous arm and tEndo Knock-in vector II had 1.06 kb and 1.8 kb homologous arm. To express endostatin gene as transgene, the F2A sequence was fused to the 5' terminal of endostatin gene and inserted into exon 7 of the ${\beta}-casein$ gene. The knock-in vector and TALEN were introduced into the bovine fibroblast by electroporation. The knock-in efficiencies of cEndo, tEndo I, and tEndo II vector were 4.6%, 2.2% and 4.8%, respectively. These results indicated that size of 3' arm in the knock-in vector is important for TALEN-mediated homologous recombination in the fibroblast. In conclusion, our knock-in system may help to create transgenic dairy cattle expressing human endostatin protein via the endogenous expression system of the bovine ${\beta}-casein$ gene in the mammary gland.

근세포 분화에 관한 연구: 차별 혼성화 스크리닝법에 의한 근원세포 분화 관련 유전자의 클로닝 (Studies on the differentiation of Myoblasts: Molecular Cloning of differentiation related Genes in the Chick Embryonic Myoblasts by Differential Hybridization.)

  • 강봉석;장세헌유병제양재섭
    • 한국동물학회지
    • /
    • 제37권2호
    • /
    • pp.240-248
    • /
    • 1994
  • 골격근 세포는 미분화 단핵 근원세포로부터 신장과 융합을 거쳐 다핵 횡문근섬유로 분화되어 가며 동시에 근특이 유전자의 발현이 선택적으로 일어난다. 본 연구에서는 계배 배양 근원세포의 분화동안 유전자 발현 조절 양상에 대한 연구를 위해, 계배 근원세포를 72시간 배양한 근섬유로부터 CDNA 라이브러리를 제작하였다. 이 cDNA 라이브러리를 미분화 단핵 근원세포(배양 36시간)와 분화된 다핵 근섬유(배양 72시간)의 poly(A)+ RNA 주형에서 합성된 [32P〕cDNA를 Probe로 사용한 differential plaque hybridization 방법으로 스크리닝하였다 분화된 다핵 근섬유 CDNA probe에 강한게 흔성화되는 CDNA clone을 선별하여 클로닝하였다. 선별한 CDNA clone 들 중 하나는 약 1.3 Kb 크기의 삽입절편을 갖고 있는 것으로 나타났고, 이 CDNA를 probe로 사용하여 northern blotting 한 결과, 이 CDNA엑 대한 유전자는 미분화 단핵 근원세포에서 분화된 다핵 근섬유로 분화가 진행됨에 따라 유전자 산물인 RNA 양이 증가되는 것으로 나타났다 또한 이 1.3 Kb CDNA에 대한 RNA의 크기는 약 2 7 Kb로 확인되었다.

  • PDF

Single-Chip Microprocessor Control for Switched Reluctance Motor Drive

  • Hao Chen;Ahn, Jin-Woo
    • KIEE International Transaction on Electrical Machinery and Energy Conversion Systems
    • /
    • 제2B권4호
    • /
    • pp.207-213
    • /
    • 2002
  • The paper introduces a switched reluctance motor drive system based on an 80C31 and an Intel 80C 196KB single-chip microprocessor control. Advance schemes are used in turn-on and turn-off angles with the power converter's main switches during traction and regenerative braking. The principles of traction speed control and braking torque control are given. The hardware and software patterns in the 80c31 and the Intel 80C196KB single-chip microprocessor control system are also presented.

투각섬석-처마카이트 각섬석 고용체의 고압합성 및 화학적 성질 (High-Pressure Synthesis and Chemistry of Amphibole Solid Solutions along the Joint Tremolite-Tschermakite)

  • 조문섭
    • 한국광물학회지
    • /
    • 제3권1호
    • /
    • pp.1-6
    • /
    • 1990
  • Clacic amphiboles along the tremolite (Tr)-tschermakite(Ts) joint were synthesized using a piston-cylinder appratus. At 750-85$0^{\circ}C$and 12-2 kb, amphibole+corundum coexist with zoisite($\pm$talc, chlorite, and Mg-staurolite), but with anorthite($\pm$cholorite, spinel, pyroxenes, and sapphirine) at lower P. At 90$0^{\circ}C$, amphibole+corundum+clinopyroxene($\pm$anorthite, forsterite, sapphirine, and garnet) are stable over the P range 12-18 kb. These amphibole-bearing assemblages are replaced at high P by clcinopyroxene+talc+chlorite+zoisite at 650-75$0^{\circ}C$, and at higher temperatures by garnet+clinopyroxene($\pm$zoisite, orthopyroxene, and Mg-staurolite). Synthetic amphiboles with Ts>~45 mol% contain as much as 0.15 excess cations per formula unit(pfu) based on 23 oxygens(anhydrous formula), whereas less tschermakitic ones are deficient in cation occupancy by up to 0.18 pfu. This trend is attributed to the 야/trioctahedral substitution in Ca-amphiboles. Compositions of synthetic amphiboles display systematic changes with P and T governed by coexisting mineral assemblages. The Ts content (=[8-Si-Na]/2) increases with increasing T( Ts/ T=~0.1 nik% K-1) in the range 750-85$0^{\circ}C$, but remains nearly constant at 850-90$0^{\circ}C$. Pressure dramatically affects the Ts content of Ca-amphiboles:it increases with P at 8-12 kb( Ts/ T=2-3 mol% K-1), but significantly decreases at 12-21 kb( Ts/ P=-2.5 mol% Kb-1). Hence, the most tschermakitic amphiboles, containing 60$\pm$5 mol % Ts, or 1.2$\pm$0.1 tetrahedral Al, occur at 12 kb and 850-90$0^{\circ}C$. Compositions of Ca-amphiboles defined by a simple reaction, 3 Tr+2 zoisite+7 corundum+H2O=5 Ts, are reversed and used to estimate thermodynamic parameters of tschermakite assuming ideal mixing of Tr-Ts solid solutions. Predicted standard molal entropy and enthalpy of tschermakite are : S$^{\circ}$of Tr-Ts solid solutions. Predicted standard molal entropy and enthalpy of tschermakite are : S$^{\circ}$=566.9$\pm$13.7 J mol-1K, -1and H$^{\circ}$=-12518.36$\pm$15.17 kJ mol.-1

  • PDF

한국 및 일본산 참굴 (Crassostrea gigas Thunberg)과 한국산 바위굴(C. nippona Seki) 의 미토콘드리아 DNA 변이 (Mitochondrial DNA Variation in Oysters (Crassostrea gigas Thunberg and C. nippona Seki) Populations from Korea and Japan)

  • 박미선;김상해
    • Animal Systematics, Evolution and Diversity
    • /
    • 제11권2호
    • /
    • pp.235-242
    • /
    • 1995
  • 한국 두 지역의 참굴(CRassostrea gigas Thunberg) 과 일본산 참굴, 그리고 한국산 바위굴(Crassostrea nippona Seki)의 유전적 근연관계를 조사하기 위하여 미토콘드리아 DNA 절편분석을 하였다. 참굴의 전체 미토콘드리아 DNA 크기는 세 지역 모두 약 18 kb 로서 동일하였으나 한국 동해산 바위굴의 경우는 약 22 kb 정도의 크기를 나타내었다. 여섯 개 염기쌍을 인식하는 8종류의 제한효소를 사용하여 분석한 결과 세 지역 참굴의 mtDNA에서 BamHI과 Bg1I 그리고 XhoI 절단시 동일한 DNA 절편이 나타나는 특징이 있었다. 종내 지역간 염기서열 치환율 (p) 은 남해안과 서해안산 참굴에서 2%로 가장 가까운 근연관계를 보였으며, 이들과 일본산 참굴 사이에는 5%의 p값을 보였다. 참굴과 바위굴, 두 종간에서는 약 42% 의 염기서열 치환율을 갖는 근연관계를 나타내었다.

  • PDF

고려인삼(Panax ginseng C.A. Meyer) ATPase $\alpha$-subunit 유전자의 Cloning (Molecular Cloning of ATPase $\alpha$-Subunit Gene from Mitochondria of Korean Ginseng (Panu ginseng C.A. Meyer))

  • Park, Ui-Sun;Choi, Kwan-Sam;Kim, Kab-Sig;Kim, Nam-Won;Choi, Kwang-Tae
    • Journal of Ginseng Research
    • /
    • 제19권1호
    • /
    • pp.56-61
    • /
    • 1995
  • Molecular cloning and restriction mapping on ATPase $\alpha$-subunit gene (atpA) were carried out to obtain genomic information concerned with the gene structure and organization in Korean ginseng mitochondria. Two different clones containing the homologous sequence of atpA gene were selected from SalI and PstI libraries of mitochondrial DNA (mtDNA) of Korean ginseng. The sizes of mtDNA fragments inserted in SalI and PstI clones were 3.4 kb and 13 kb, respectively. Southern blot analysis with [$^{32}P$] labelled Oenothera atPA gene probe showed that atpA gene sequence was located in 2.0 kb XkaI fragment in PstI clone and in 1.7 kb XbaI fragment in SalI clone. A partial sequening ascertained that the SalI clone included about 1.2 kb fragment from SalI restriction site to C-terminal sequence of this gene but about 0.3 kb N-terminal sequence of open reading frame was abscent. The PstI fragment was enough large to cover the full sequence of atpA gene. The same restriction pattern of the overlapped region suggests that both clones include the same fragment of atiA locus. Data of Southern blot analysis and partial nucleotide sequencing suggested that mtDNA of Korean ginseng has a single copy of atpA gene. Key words ATPase a-subunit, mitochondrial DNA, Panax ginseng.

  • PDF

Saccharomyces cerevisiae에서 RNA1 유전자의 클로닝 (Cloning of RNA1 Gene from Saccharomyces cerevisiae)

  • 송영환;고상석;이영석;강현삼
    • 미생물학회지
    • /
    • 제27권2호
    • /
    • pp.77-84
    • /
    • 1989
  • Saccharomyces cerevisiae의 RNAI 유전자의 온도 감수성 돌연변이 균주는 성장허용 온도인 23"C에서는 정상적인 성"J--을하나, 성 장억제 온도인 36°C에서는 tRNA, rRNA 그러고 mRNA 의 선우울질을을 핵내에 축적함으후써 성장을 못한다. 본 실 험에서는 complementation 에 의하여 RNAI 유전자를 클로녕하였으며 concomitant loss 실험에 의하여 이 유천자의 클로닝 을 확인하였다. 유전자의 위치를 확인한 결과 3.5kb의 Bgl II 조각내에 RNAI 유전자가 존재함을 알 수 있였으며, 2.1kb에 해당하는 BamH I -Bgl II 조각에서는 RNAI 유전지에 의한 complementation 능력이 상실되는 것으후 보아 RNAI 유전자 는 3.5kb의 BglIl 조각내에 포함되는 BamH 1 자리 주위에 결쳐 있픔을 알 수 있었다.

  • PDF

해면의 화학적 성분 연구 (The Chemical Constituents from Unidentified Sponge)

  • 박선구
    • 대한화학회지
    • /
    • 제38권2호
    • /
    • pp.169-173
    • /
    • 1994
  • 인도네시아 Manado만의 Sulawesi에서 채집한 미동정된 해면으로부터 KB cancer cell line에 대해 활성을 갖는 xestoquinone, halenaquinol sulfate 및 halenaquinol이 검출되었다. 이들을 $^1H-,\;^{13}C$-NMR, $^1H-,\;^{13}C$(1 bond) heteronuclear Multiple Quantum Cogerence Spectroscopy$(HMQC)^1$, $^1H-,\;^{13}C$C(2 and 3 bond) Heteronuclear multiple Bond Correlation Spectroscopy$(HMBC)^2$, Electron Impact Mass Spectroscopy(EI ms), UV 및 IR에 의해 밝혔다.

  • PDF

Comparative Proteome Analysis of Two Antagonist Bacillus subtilis Strains

  • Zhang, C.X.;Zhao, X.;Han, F.;Yang, M.F.;Chen, H.;Chida, T.;Shen, S.H.
    • Journal of Microbiology and Biotechnology
    • /
    • 제19권4호
    • /
    • pp.351-357
    • /
    • 2009
  • Natural wild-type strains of Bacillus subtilis are extensively used in agriculture as biocontrol agents for plants. This study examined two antagonist B. subtilis strains, KB-1111 and KB-1122, and the results illustrated that KB-1122 was a more potent inhibitor of the indicator pathogen than KB-1111. Thus, to investigate the intrinsic differences between the two antagonist strains under normal culture conditions, samples of KB-1111 and KB-1122 were analyzed using MALDI-TOF-MS. The main differences were related to 20 abundant intracellular and 17 extracellular proteins. When searching the NCBI database, a number of the differentially expressed proteins were identified, including 11 cellular proteins and 10 secretory proteins. Among these proteins, class III stress-response-related ATPase, aconitate hydratase, alpha-amylase precursor, and a secretory protein, endo-l, 4-beta-glucanase, were differentially expressed by the two strains. These results are useful to comprehend the intrinsic differences between the antagonism of KB-1111 and KB-1122.

Thermodynamic Evaluation of Fe Effect on Alumina Solubility in Orthopyroxene coexisting with Garnet

  • Lee, Han-Yeang
    • 한국지구과학회지
    • /
    • 제25권1호
    • /
    • pp.39-47
    • /
    • 2004
  • In the FMAS system the effect of iron on alumina solubility in orthopyroxene has been determined by experiments with crystalline starting mixtures of garnet and orthopyroxene of known initial compositions at 20 kb, 975$^{\circ}C$ and 25 kb, 1,200$^{\circ}C$. These data have been modeled to develop a thermodynamic method for the calculation of Al$_{2}$O$_{3}$ in orthopyroxene as a function of P, T and composition. The direct application of the alumina solubility data in the MAS system to natural assemblages could lead to significant overestimation of pressure, probably by about 5kb for the relatively common garnet-lherzolites with abot 25 mol per cent Ca + Fe$^{2+}$ in garnet and about 1 wt. per cent Al$_{2}$O$_{3}$ in orthopyroxene since the effect of Fe is similar to that of Ca and Cr$^{3+}$in reducing the alumina solubility in orthopyroxene in equilibrum with garnet relative to that in the MAS system.