• 제목/요약/키워드: co-transformation

검색결과 673건 처리시간 0.025초

Development of Transgenic Orchardgrass (Dactylis glomerata L.) Using Agrobacterium tumefaciens

  • Lee, Hyoshin;Lee, Byung-Hyun;Jinki Jo
    • 한국초지조사료학회지
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    • 제20권2호
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    • pp.103-108
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    • 2000
  • To produce of transgenic orchardgrass, the seed-derived calli of orchardgrass (Dactylis glomerata L.) co-cultivated with Agrobacterium turnefaciens EHAlOl harboring binary vector pIG121-Hm were selected with hygromycin and then transferred onto N6 regeneration medium containing 1 rngl l of NAA, 5 rngl l of kinetin, 250 rngl l of carbenicillin and 50 mg/ l of hygromycin. The efficiency of transformation was differed on cultivars, that is, 'Potomac' appeared 12% of transformation efficiency while 'Amba' did 5.5%. The addition of acetosyringone during co-cultivation was a key to successhl transformation of orchardgrass. Transgene fragments were identified by PCR analysis and the constitutive expression of GUS gene was confirmed by Northern blot analysis. (Key words : Acetosyringone, Agrobacterium tumefaciens, Orchardgrass (Dactylis glomerata L.), Transformation)

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저온상변태법을 이용한 주석 및 산화주석 나노말의 제조 (Fabrication of Sn and SnO2 Nanopowders by Low-Temperature Phase Transformation Method)

  • 이근재;주연준;소용대;김남훈;이재성;좌용호
    • 한국분말재료학회지
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    • 제13권1호
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    • pp.46-51
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    • 2006
  • Through the volume change of Sn in a low-temperature phase transformation, the Sn nanopowder with high, purity, was fabricated by an economic and eco-friendly process. The fine cracks were spontaneously generated. in, Sn ingot, which was reduced to powders in the repetition of phase transformation. The Sn nanopowder with 50 run in size was obtained by the 24th repetitions of phase transformation by low-temperature and ultrasonic treatments. Also, the $SnO_2$ powder was fabricated by the oxidation of the produced Sn powder to the ingot and milled by the ultrasonic milling method. The $SnO_2$ nanopowder of 20 nm in size was fabricated after the milling for 180 h.

Insertion Mutation in HMG-CoA Lyase Increases the Production Yield of MPA through Agrobacterium tumefaciens-Mediated Transformation

  • Dong, Yuguo;Zhang, Jian;Xu, Rui;Lv, Xinxin;Wang, Lihua;Sun, Aiyou;Wei, Dongzhi
    • Journal of Microbiology and Biotechnology
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    • 제26권11호
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    • pp.1924-1932
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    • 2016
  • Mycophenolic acid (MPA) is an antibiotic produced by Penicillium brevicompactum. MPA has antifungal, antineoplastic, and immunosuppressive functions, among others. ${\beta}-Hydroxy-{\beta}-methylglutaryl-CoA$ (HMG-CoA) lyase is a key enzyme in the bypass metabolic pathway. The inhibitory activity of HMG-CoA lyase increases the MPA biosynthetic flux by reducing the generation of by-products. In this study, we cloned the P. brevicompactum HMG-CoA lyase gene using the thermal asymmetric interlaced polymerase chain reaction and gene walking technology. Agrobacterium tumefaciens-mediated transformation (ATMT) was used to insert a mutated HMG-CoA lyase gene into P. brevicompactum. Successful insertion of the HMG-CoA lyase gene was confirmed by hygromycin screening, PCR, Southern blot analysis, and enzyme content assay. The maximum MPA production by transformants was 2.94 g/l. This was 71% higher than wild-type ATCC 16024. Our results demonstrate that ATMT may be an alternative practical genetic tool for directional transformation of P. brevicompactum.

Development of Transgenic Tall Fescue Plants from Mature Seed-derived Callus via Agrobacterium-mediated Transformation

  • Lee, Sang-Hoon;Lee, Dong-Gi;Woo, Hyun-Sook;Lee, Byung-Hyun
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권10호
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    • pp.1390-1394
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    • 2004
  • We have achieved efficient transformation system for forage-type tall fescue plants by Agrobacterium tumefaciens. Mature seed-derived embryogenic calli were infected and co-cultivated with each of three A. tumefaciens strains, all of which harbored a standard binary vector pIG121Hm encoding the neomycin phosphotransferase II (NPTII), hygromycin phosphotransferase (HPT) and intron-containing $\beta$-glucuronidase (intron-GUS) genes in the T-DNA region. Transformation efficiency was influenced by the A. tumefaciens strain, addition of the phenolic compound acetosyringone and duration of vacuum treatment. Of the three A. tumefaciens strains tested, EHA101/pIG121Hm was found to be most effective followed by GV3101/pIG121Hm and LBA4404/pIG121Hm for transient GUS expression after 3 days co-cultivation. Inclusion of 100 $\mu$M acetosyringone in both the inoculation and co-cultivation media lead to an improvement in transient GUS expression observed in targeted calli. Vacuum treatment during infection of calli with A. tumefaciens strains increased transformation efficiency. The highest stable transformation efficiency of transgenic plants was obtained when mature seed-derived calli infected with A. tumefaciens EHA101/pIG121Hm in the presence of 100 $\mu$M acetosyringone and vacuum treatment for 30 min. Southern blot analysis indicated integration of the transgene into the genome of tall fescue. The transformation system developed in this study would be useful for Agrobacterium-mediated genetic transformation of tall fescue plants with genes of agronomic importance.

캘러스 유도에 의한 수박 형질전환 (Genetic Transformation of Watermelon (Citrullus vulgaris Schard.) by Callus Induction)

  • 권정희;박상미;임미영;신윤섭;한지학
    • Journal of Plant Biotechnology
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    • 제34권1호
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    • pp.37-45
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    • 2007
  • 수박은 형질전환이 매우 어렵고 직접적인 신초 유도방법으로는 안정된 형질전환을 기대할 수 없어서, 다른 여러 작물에서 높은 효율을 보였던 캘러스 유래 신초 유도 방법을 도입하고자 하였다. 수박의 최적 캘러스 유도조건은 자엽 절편체의 경우 2.0 mg/L zeatin과 0.1 mg/L IAA이었으며 뿌리 절편체의 경우 2.0 mg/L BA와 0.1 mg/L 2,4-D이었다. NptII 유전자의 선발 항생제는 kanamycin보다는 paromomycin 빠르고 효과적이었으며, 수박의 절편체에 Agrobacterium을 접종 한 후 paromomycin을 125 mg/L 첨가한 배지에서 선발하였다. pmGFP5-ER vector로 형질전환한 후 캘러스 상태에서 형광현미경을 통해 GFP 유전자의 도입을 확인하였으며, 딱딱한 초록색의 캘러스에서 강한 GFP 발현을 관찰하였고, 자엽유래 캘러스의 경우 WM8에서 9.0%, 뿌리유래 캘러스의 경우 WM6에서 8.3%의 가장 높은 GFP 발현 효율을 보였다. GFP 유전자 도입과 같은 방법으로 WMV-CP 유전자가 있는 pWMV2300 vector로 형질전환한 후 캘러스 상태에서 PCR 및 Southern blot 분석을 한 결과, 두 점의 캘러스에서 WMV-CP 유전자가 도입되었음을 확인하였다. 본 연구를 통하여 확립된 수박의 캘러스 유도 시스템은 안정된 수박 형질전환 방법에 기초 자료로서 이용될 것이다.

Co-sputtering에 의해 증착된 TiNi 박막의 미세조직 및 기계적성질에 미치는 어닐링 열처리 효과 (Effects of Annealing Treatments on Microstructure and Mechanical Property of co-sputtered TiNi Thin Film)

  • 박수대;백창현;홍주화
    • 열처리공학회지
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    • 제21권1호
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    • pp.26-32
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    • 2008
  • Effects of annealing treatment on microstructure and mechanical property of co-sputtered TiNi thin films were studied. As-deposited films showed amorphous state. However, above annealing temperature of $500^{\circ}C$ martensite phase (B19'), precipitate phase ($Ti_2Ni$) and a small amount of parent phase ($B_2$) were present, and phase transformation behaviors were three multi-step phase transformations $B19^{\prime}{\rightarrow}B_2$ and $B_2{\rightarrow}R-phase$ and $R-phase{\rightarrow}B19^{\prime}$. Increase of martensite transformation temperature, increase of microhardness and Young's modulus of TiNi films annealed above $500^{\circ}C$ were discussed in terms of precipitate phase.

Efficiency of transformation mediated by Agrobacterium tumefaciens using vacuum infiltration in rice (Oryza sativa L.)

  • Safitri, Fika Ayu;Ubaidillah, Mohammad;Kim, Kyung-Min
    • Journal of Plant Biotechnology
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    • 제43권1호
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    • pp.66-75
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    • 2016
  • Agrobacterium-mediated gene transfer has recently been developed to improve rice transformation. In this study, 3 different transformation methods were tested including soaking, co-cultivation, and vacuum infiltration. Agrobacterium tumefaciens GV3101 harboring the binary vector pGreen:: LeGSNOR was used in this experiment. This study aimed to identify the most appropriate method for transferring LeGSNOR into rice. Vacuum infiltration of the embryonic calli for 5 min in Ilpum resulted in high transformation efficiency based on confirmation by PCR, RT-PCR, and qRT-PCR analyses. In conclusion, we described the development of an efficient transformation protocol for the stable integration of foreign genes into rice; furthermore, the study results confirmed that PCR is suitable for efficient detection of the integrated gene. The vacuum infiltration system is a potentially useful tool for future studies focusing on transferring important genes into rice seed calli, and may help reduce time and effort.

Co-registration of Multiple Postmortem Brain Slices to Corresponding MRIs Using Voxel Similarity Measures and Slice-to-Volume Transformation

  • Kim Tae-Seong
    • 대한의용생체공학회:의공학회지
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    • 제26권4호
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    • pp.231-241
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    • 2005
  • New methods to register multiple hemispheric slices of the postmortem brain to anatomically corresponding in-vivo MRI slices within a 3D volumetric MRI are presented. Gel-embedding and fiducial markers are used to reduce geometrical distortions in the postmortem brain volume. The registration algorithm relies on a recursive extraction of warped MRI slices from the reference MRI volume using a modified non-linear polynomial transformation until matching slices are found. Eight different voxel similarity measures are tested to get the best co-registration cost and the results show that combination of two different similarity measures shows the best performance. After validating the implementation and approach through simulation studies, the presented methods are applied to real data. The results demonstrate the feasibility and practicability of the presented co­registration methods, thus providing a means of MR signal analysis and histological examination of tissue lesions via co­registered images of postmortem brain slices and their corresponding MRI sections. With this approach, it is possible to investigate the pathology of a disease through both routinely acquired MRls and postmortem brain slices, thus improving the understanding of the pathological substrates and their progression.

레이저 박리용 선형 빔 광학계의 빔 변환 모듈과 단축 형상에 대한 연구 (Research on BTU and Short-axis Geometry of Line-beam Optics for LLO Applications)

  • 이승민;이광진;김대용;이상현;정진호
    • 한국광학회지
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    • 제32권6호
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    • pp.276-285
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    • 2021
  • 본 논문은 디스플레이용 OLED의 제작 공정에 사용하는 레이저 박리(laser lift-off, LLO) 장비의 선형빔 광학계에 대한 연구로서, M2가 큰 레이저를 사용하는 경우, 단축의 선폭을 작게 만들면서 공정심도를 깊게 유지하기 위한 빔 변환 모듈(beam transformation unit, BTU)을 포함한 광학계에 대한 연구를 진행하였고, 단축 형상을 슈퍼 가우시안으로 만들 수 있는 방법과 flat-top 분포로 만들 수 있는 광섬유를 사용하는 빔 변환 방식에 대하여 제시하였다.

Na2CO3의 첨가에 따른 석탄 선탄 폐석의 소결 상변화 연구 (Phase Transformation of Coal Tailing of Beneficiation with the Addition of Na2CO3 at High Temperature)

  • 유광석
    • 자원리싸이클링
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    • 제29권6호
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    • pp.73-78
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    • 2020
  • 선탄 폐석의 유리 소재 원료로의 활용을 위해 선탄 폐석의 고온 상 전위 거동 및 Na2CO3의 첨가에 따른 유리질(비정질)화 실험을 실시하였다. 실험 결과 700~1,100 ℃에서 소성된 선탄폐석에는 mullite가 생성되고, 1,450 ℃에서는 mullite와 cristobalite만이 존재하였다. 선탄 폐석의 유리질화는 Na2CO3 첨가됨에 따라 진행되었으며, Na2CO3 10 wt.% 첨가 후 1,450 ℃ 온도에서 소성된 선탄 폐석의 유리질화는 97.9 wt.%로 나타났다. 그러나 Na2CO3가 20 wt.% 첨가된 선탄 폐석의 시료의 경우, 1,100 ℃에서의 2차 재소성 과정에서 nepheline(Na2O·Al2O3·2SiO2)이 생성되었으며, 이를 통해 선탄폐석의 유리질화를 위한 최적의 Na2CO3 첨가량은 10 wt.%로 나타났다.