• Title/Summary/Keyword: chimeric

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Expression of Mouse Adenosine Deaminase Gene in Transgenic Tobacco (Nicotiana tabacum L.) (형질전환 연초(Nicotiana tabacum L.)의 Mouse Adenosine Deaminase 유전자 발현)

  • 양덕춘;박지창;최광태;이정명
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.4
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    • pp.195-200
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    • 1995
  • The mammalian adenosine deaminase(ADA) gene was stably expressed in transgenic tobacco plane. The chimeric ADA gene 35S/35S/AMV/ADA/Tnos, has been constructed. This chimeric gene was introduced into the binary vector pRD400, which was thereafter mobilized into Agrobacterium tumefaiens strain MP90 harboring disarmed Ti-plasmid. The resulting strains were used to transform Nicofiana tabacum L. using the leaf disc. Incorporation of the chimeric gene into plant were confirmed by PCR and Northern blot analyses. Immunoblot analysis showed that ADA protein was successfully synthesized in the transgenic tobacco plants.

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Transfer of R plasmids of Bacterial Isolates and Their Cloned R Genes in Natural Wastewater Environments (I) -Cloning of $Km^rCm^r$Gene- (하폐수의 자연환경에서 R plasmid와 재조합 유전자의 전이특성( I ) -$Km^rCm^r$유전자의 클로닝-)

  • 김치경;이성기
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.447-453
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    • 1989
  • In order to study the transfer of antibiotics resistance genes of the genetically cloned bacteria in water environments, DK1 strain, which is resistant to kanamycin (Km), chloramphenicol (Cm), streptomycin (Sm), and sulfadiazine (Su), was selected from the Gram-negative bacterial isolates from wastewater. One of 4 plasmids harboured in the DK1 strain was found to possess Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ gene and be about 68 kb in size, and it was designated as pDK101. The plasmid of pDK101 was also found to have 16, 32, and 6 restriction sites for EcoRI. .PstI, and SalI, respectively. From the digestion fragments of pDK101 plasmid and pKT230 used as a vector by EcoRI restriction endonuclease, pDT309 and pDT529 were constructed as chimeric plasmids which possess Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ gene and are 30.9 and 52.9 kb in size, respectively. When the chimeric plasmids were trasformed into E. coli C600 or E. coli HB101, transformants of DKC601, DKC602, DKH102, and DKH103 were obtained as cloned bacterial cells. The Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ genes were well expressed in those cloned cells and the chimeric plasmids were clearly detected in the cloned cells of DKC601 and DKH103.

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Analysis of the Glycinin Gy2 Promoter Activity in Soybean Protoplasts and Transgenic Tobacco Plants (대두 원형질체와 형질전환된 담배에서의 대두 glycinin 유전자 Gy2 promoter의 발현조절 기작)

  • Kim, Soo-Jung;Lee, Jee-Young;Kim, Chung-Ho;Choi, Yang-Do
    • Applied Biological Chemistry
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    • v.38 no.5
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    • pp.387-392
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    • 1995
  • To study the regulatory expression mechanism of soybean glycinin gone, Gy2, the 5' upstream region of the gene was searched for the presence of putative regulatory elements by nucleotide sequencing. It revealed various kinds of regulatory sequence elements commonly found in plant storage protein genes. There were canonical promoter sequences, TATA box (TATAAT) and AGGA box (GAAT) which are common in the 5' upstream region of the plant genes. The embryo factor binding sequence, RY repeat, CACA sequences, ${\alpha}$-conglycinin enhancer-like sequences were also found. To delineate the function of these sequences, 5' upstream deletion mutants of Gy2 were prepared and fused to the ${\alpha}$-glucuronidase (GUS) gene. Each chimeric construct was transferred into soybean protoplasts for transient assay, which led to the identification of the sequences between -281 and -223, -170 and -122, of Gy2 promoter as negative regulatory elements, and the sequences between -223 and -170, -122 and -16 as positive regulatory elements. These results are consistent in transformed tobacco plants as well. The serially deleted promoter fragments fused to the GUS were transformed into Nicotiana tabacum by Agrobacterium tumefaciens using the binary vector system. GUS activity of Gy2 promoter deletion constructs was detected only in seeds but not in leaves with different levels of expression as in transient assay. These results suggest that the glycinin Gy2 promoter drives a tissue-specific expression in transgenic tobacco plants.

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Expression of Chitinase Gene in Solanum tuberosum L.

  • Park, Kyung-Hwa;Yang, Deok-Chun;Jeon, Jae-Heung;Kim, Hyun-Soon;Joung, Young-Hee;Hyouk Joung
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.85-90
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    • 1999
  • In order to protect fungal diseases, leaf disc explants of Solanum tuberosum cultivar, Belchip, was infected with an Agrobacterium MP90 strain containing chimeric gene construct, consisting of antibiotic resistance and chitinase gene driven by the CaMV 35S promoter, for transformation. Regenerated multiple shoots were selected on a medium containing kanamycin and carbenicillin after exposure to Agrobacterium. The presence and integration of the npt II and chitinase gene were confirmed by polymerase chain reaction(PCR). Northern blot analysis indicated that the genes coding for the enzyme could be expressed in potato plants. The chitinase activity of transgenic potato plants was higher than the control potato.

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Developmental potential of bisected-aggregated mouse embryos after freezing (동결보존한 마우스 이분집합배의 생존에 관한 연구)

  • Shin, Sang-tae;Jo, Choong-ho
    • Korean Journal of Veterinary Research
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    • v.31 no.2
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    • pp.229-234
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    • 1991
  • Thc chimeric morulae were produced following aggregation of the half embryos which were microsurgically bisected at 8-cell and early morula stage. Different phenotypic embryos were obtained by mating ICR female mice with ICR or CBA male mice. The early morula stage was thc desirable stage for the aggregation of mouse embryos after bisection. The post-thawed survival rates of bisected-aggregated embryos that developed into normal blastocyst after conventional freezing in DMSO and ethylene glycol were 30.5 and 32.896, respectively. One offspring was produced by transferring the 67 frozen-thawed bisected-aggregated embryos.

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Effect of Heparin on the High Affinity KGF and aFGF Binding to the Chimeric KGFR-HFc

  • Cheon, Hyae-Gyeong
    • BMB Reports
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    • v.29 no.3
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    • pp.205-209
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    • 1996
  • To investigate the role of heparin in keratinocyte growth factor (KGF) and acidic fibroblast growth factor (aFGF) high affinity binding to the KGF receptor (KGFR), a cell free system was established which utilized a secreted chimeric molecule between the KGFR extracellular domain and the immunoglobulin heavy chain Fc domain (KGFR-HFc). KGFR-HFc was purified from NIH 3T3 cells and demonstrated the binding of $[^3H]-heparin$ as well as heparin Sepharose. Scatchard analysis showed that the dissociation constant for heparin binding to KGFR-HFc was 140 nM. High affinity KGF and aFGF binding to KGFR-HFc remained unchanged after treatment with 0.6 M NaCl, which is the concentration sufficient to release any bound heparin to the KGFR-HFc. These results strongly suggest that although the KGFR interacts with heparin, the presence of heparin is not absolutely required for high affinity binding of either KGF or aFGF to the KGFR.

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Unplanned change from double free flap to a chimeric anterolateral thigh flap in recurrent laryngeal cancer

  • Ki, Sae Hwi;Ma, Sung Hwan;Sim, Seung Hyun;Choi, Matthew Seung Suk
    • Archives of Craniofacial Surgery
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    • v.20 no.6
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    • pp.416-420
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    • 2019
  • Reconstruction method choice in recurrent head and neck cancer depends on surgical history, radiation therapy dosage, conditions of recipient vessels, and general patient condition. Furthermore, when defects are multiple or three dimensional in nature, reconstruction and flap choice aimed at rebuilding the functional structure of the head and neck are difficult. We experienced successful reconstruction of recurrent laryngeal cancer requiring reconstruction of esophageal and tracheostomy stroma defects using a chimeric two-skin anterolateral thigh flap with a single pedicle.

Tobacco plant transformed with a coat protein gene sequence of TMV (TMV외피 단백질 유전자의 연초로의 형질전환)

  • 이기원;박성원;김남원;박은경
    • Journal of the Korean Society of Tobacco Science
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    • v.15 no.2
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    • pp.161-166
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    • 1993
  • A double - stranded cDNA fragment (436bp) encoding coat protein of tobacco mosaic virus(TMV) was derived from the total 480nucleotides gene after reverse transcription of TMV RNA, and subclorled into a plant expression vector pBl 121, resulting in pBL 430. The plasmid DNA containing this chimeric gene was moved from E. cofi to Agrobacterium tumefaciens strain A28l, and was introduced in시 the tobacco plant by the Agrobocterium Ti - mediated transformtion system. The transformants were selected on a selection media containing kanamycin. The shoots add roots could be differentiated from the explants and whole plants were obtained. From Southern blot hybridization analysis, DNA extracted from transformants, it could be conformed that the chimeric gene fragment was inserted into the genomic DNA of tobacco plant.

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