• Title/Summary/Keyword: DISC-virus

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Isolation of Coat Protein Gene from Cucumber Mosaic Virus and Its Introduction into Tobacco (오이 모자이크 바이러스 외피 단백질 유전자 분리 및 담배로의 형질전환)

  • 손성한;김경환;김영태;박종석;김주곤;이광웅;황영수
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.149-155
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    • 1995
  • The coat protein (CP) gene was cloned from RNA genome of the Cucumber Mosaic Virus strain ABI (CMV-ABI) isolated in Korea. The comparisons of the nucleotide sequence of the cloned CP gene and its deduced amino acid sequences with other CP genes revealed that the CMV-ABI belongs to subgroup I (type I), CMV-ABI developed the typical mosaic symptom in infected plants. Tobacco plants (Samsun and NC82) were transformed by leaf-disc transformation via Agrobacterium, temefaciens LB4404 harboring pVCP, witch CMV-ABI CP gene was inserted into the pBI121, and a number of mature transgenic tobacco plants were developed. Southern and PCR analysis of genomic DNA from the transgenic plants showed that the CP gene was integrated into the genomes of the most of the transgenic plant. Result of the segregation patterns of resistance in T1 seedlings of the plants to kanamycin showed that the transgenic plants containing l,2 and 3 copies of CP gene were50%, 39% and 11% of the total transgenic plants, respectively.

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Introduction of VP6 Gene into Potato Plant by Agrobacterium-mediated Transformation and Analysis of VP6 Expression in Transgenic Potatoes (Rotavirus VP6 유전자의 감자식물체내로의 도입과 형질전환체의 발현분석)

  • Youm, Jung-Won;Jeon, Jae-Heung;Jung, Jae-Yeol;Lee, Byoung-Chan;Kang, Won-Jin;Kim, Mi-Sun;Kim, Chul-Joong;Joung, Hyouk;Kim, Hyun-Soon
    • Journal of Plant Biotechnology
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    • v.29 no.2
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    • pp.93-98
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    • 2002
  • A VP6 fragments was subcloned with BamHI in the binary pMBP-1 vector under Califlower Mosaic Virus (CaMV) 355 promoter and neomycin phosphotransferase II (npt II) gene. The recombinant binary vector was mobilized into Agrobacterium-tumefaciens LBA4404 by the freeze-thaw method and potato (Solanum tubensum L. cv Desiree) was transformed by modified leaf-disc cocultivation. Shoots were induced on MS medium with 0.01 mg/L NAA, 0.1 mg/L GA$_3$, 2.0 mg/L Zeatin, 100.0 mg/L kanamycin, 500.0 mg/L carbenicillin. In order to identify the copy number of VP6 into potato plant, total genomic DNA was isolated from transgenic potato and analysed by Southern blotting. Genomic DNA and total mRNA analysis demonstrated the incorporation of the foreign gene into the potato genome, as well as their transcription.

A Study of the Retrovirus-Mediated Transgenic Chicken Production on Chicken Embryos (닭 수정란에서 Retrovirus를 이용한 형질전환 닭 생산 연구)

  • Byun S. J.;Park C.;Kim S. W.;Park J. K.;Chang W. K.;Yang B. S.;Kim T. Y.;Sohn S. H.;Kim S. H.;Jeon I. S.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.225-229
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    • 2005
  • Microinjection of recombinant retrovirus beneath the blastoderm of non-incubated chicken embryo is now the most widespread method for generating transgenic chickens, but transgenesis rates are very low. So to improve this problem, we first introduced retrovirus vector carrying RSV-GFP gene to an one-cell embryo culture system. To investigate whether retrovirus could work on an one-cell chicken embryo, we microinjected the concentrated retrovirus stocks into the germinal disc of one cell or stage-X chicken embryos. Analysis of reporter gene expression on day 4 embryos showed that GFP expression was observed in the only stage-X chicken embryo but was not in the one-cell embryo group. These results suggest that retrovirus system is the most efficient method to generate transgenic chickens in the stage-X embryo.