• Title/Summary/Keyword: transgenic maize

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Expression of Dengue virus EIII domain-coding gene in maize as an edible vaccine candidate

  • Kim, Hyun A;Kwon, Suk Yoon;Yang, Moon Sik;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.50-55
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    • 2014
  • Plant-based vaccines possess some advantages over other types of vaccine biotechnology such as safety, low cost of mass vaccination programs, and wider use of vaccines for medicine. This study was undertaken to develop the transgenic maize as edible vaccine candidates for humans. The immature embryos of HiII genotype were inoculated with A. tumefaciens strain C58C1 containing the binary vectors (V662 or V663). The vectors carrying nptII gene as selection marker and scEDIII (V662) or wCTB-scEDIII (V663) target gene, which code EIII proteins inhibite viral adsorption by cells. In total, 721 maize immature embryos were transformed and twenty-two putative transgenic plants were regenerated after 12 weeks selection regime. Of them, two- and six-plants were proved to be integrated with scEDIII and wCTB-scEDIII genes, respectively, by Southern blot analysis. However, only one plant (V662-29-3864) can express the gene of interest confirmed by Northern blot analysis. These results demonstrated that this plant could be used as a candidated source of the vaccine production.

Expression of CP4 5-Enol-Pyruvylshikimate-3- Phosphate Synthase Transgene in Inbred Line of Korean Domestic Maize (Zea may L.) (국내 옥수수 순계주에서 CP4 5-Enol- Pyruvylshikimate-3- Phosphate Synthase 유전자의 발현)

  • Cho, Mi-Ae;Kwon, Suk-Yoon;Kim, Jin-Seog;Lee, Byoung-Kyu;Moon, Choo-Yeun;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.375-380
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    • 2007
  • This study was conducted to develop herbicide-resistance domestic maize plants by introducing the CP4 5-enol-pyruvylshikimate-3-phosphate synthase (CP4 EPSPS) gene using Agrobacterium tumefaciens-mediated immature embryo transformation. Immature embryos of five genotypes (HW1, KL103, HW3, HW4, HW7) were co-cultivated with strains Agrobacterium tumefaciens (strain C58C1) containing the binary vector (pCAMBIA2300) carrying Ubiquitin promoter-CP4 EPSPS gene and Cauliflower mosaic virus 35S (CaMV35S) promoter-nptll gene conferring resistance to paromomycin as a selective agent. The presence and expression of CP4 EPSPS transgene were confirmed by PCR, RT-PCR and Northern blot analysis, respectively. Also, the resistance to glyphosate in the transgenic maize ($T_1$) was analyzed by shikimate accumulation assay. The frequency (%) of paromomycin-resistance callus was 0.37, 0.03, 2.20, 2.37, and 0.81% in pure lines HW1, KL103, HW3, HW4 and HW7, respectively. EPSP transgene sequences were amplified in putative transgenic plants that regenerated from paromomycin-resistance calli of two inbred lines (HW3, HW4). Of them, RT-PCR and Northern blot analyses revealed that the transgene was only expressed in two transgenic events (M266, M104) of HW4 inbred line, and a mild glyphosate resistance of transgenic event (M266) was confirmed by the lower shikimate accumulation in leaf segments. These results demonstrate that transgenic maize with herbicide-resistance traits in Korean genotype can be genetically obtained.

Production of Transgenic Maize (Zea mays L.) Using Agrobacterium tumefaciens-Mediated Transformation (Agrobacterium tumefaciens 공동배양법을 이용한 옥수수 형질전환체 생산)

  • Cho Mi-Ae;Park Yun-Ok;Kim Jin-Suck;Park Ki-Jin;Min Hwang-Ki;Liu Jang-Ryol;Clemente Tom;Choi Pil-Son
    • Journal of Plant Biotechnology
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    • v.32 no.2
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    • pp.91-95
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    • 2005
  • Agrobacterium tumefaciens-mediated immature embryo transformation was used to produce transgenic maize. Immature embryo of Hi II genotype were co-cultivated with strains Agrobacterium tumefaciens (C58C1) containing the binary vectors (pPTN290) carrying with Ubiquitin promoter-GUS gene as reporter gene and NOS promoter-nptll gene conferring resistance to paromomycin as selective agent. Seven embryogenic callus lines transformed showed the resistance in paromomycin antibiotics. Histochemical GUS assay showed that 7 individual lines transformed with the GUS gene were positive response among the transformants. Southern blot analysis revealed that the nptll gene segregated and expressed in their progeny.

A Safety Evaluation of Genetically Modified Feedstuffs for Livestock Production; the Fate of Transgenic DNA and Proteins

  • Beever, D.E.;Glenn, K.;Phipps, R.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.5
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    • pp.764-772
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    • 2003
  • Two genetic constructs used to confer improved agronomic characteristics, namely herbicide tolerance (HT) in maize and soyabean and insect resistance (Bt) in maize, are considered in respect of feeding to farm livestock, animal performance and the nutritional value and safety of animal products. A review of nucleic acid (DNA) and protein digestion in farm livestock concludes that the frequency of intact transgenic DNA and proteins of GM and non-GM crops being absorbed is minimal/non existent, although there is some evidence of the presence of short fragments of rubisco DNA of non-GM soya in animal tissues. It has been established that feed processing (especially heat) prior to feeding causes significant disruption of plant DNA. Studies with ruminant and non-ruminant farm livestock offered GM feeds demonstrated that animal performance and product composition are unaffected and that there is no evidence of transgenic DNA or proteins of current GM in the products of animals consuming such feeds. On this evidence, current HT and Bt constructs represent no threat to the health of animals, or humans consuming the products of such animals. However as new GM constructs become available it will be necessary to subject these to rigorous evaluation.

Identification of Excision of Ac Transposable Element in P.nigra x maximowiczii Using Agrobacterium-mediated Transformation

  • Ahn, In-Suk;Park, Young-Goo;Shin, Dong-Ill;Sul, Ill-Whan
    • Journal of Plant Biotechnology
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    • v.5 no.1
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    • pp.19-23
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    • 2003
  • The Ac (activator) which is one of the well-characterized transposable elements from maize was examined for its transposition possibility to the heterologous plant (P.nigra x maximowiczii) genome via Agrobacterium tumefacience (LBA4404) mediated transformation system. A number of transgenic plants were successfully recovered after 30 weeks by amount reduction from 50 to 15 g/$m\ell$ kanamycin for in vitro selection to minimize phytotoxic effects and to increase callus growth and regeneration efficiency. Among transgenic plants, 62 out of 106 transgenic poplars (58.5%) showed abnormal phenotypes such as severe serrated leaves and light leaf coloration. Indigo staining with X-gluc proved indirectly the restoration of Gus enzyme function and the presence of Ac in poplar genome by PCR. Southern analysis indicated the transposition and existence of Ac element in poplar genomes. In this research, an Agrobacterium-mediated transformation system in poplar species was developed and identified that Ac derived from maize can be excised and trans posed into other poplar genomes.

Expression of Indica rice OsBADH1 gene under salinity stress in transgenic tobacco

  • Hasthanasombut, Supaporn;Ntui, Valentine;Supaibulwatana, Kanyaratt;Mii, Masahiro;Nakamura, Ikuo
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.75-83
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    • 2010
  • Glycine betaine has been reported as an osmoprotectant compound conferring tolerance to salinity and osmotic stresses in plants. We previously found that the expression of betaine aldehyde dehydrogenase 1 gene (OsBADH1), encoding a key enzyme for glycine betaine biosynthesis pathway, showed close correlation with salt tolerance of rice. In this study, the expression of the OsBADH1 gene in transgenic tobacco was investigated in response to salt stress using a transgenic approach. Transgenic tobacco plants expressing the OsBADH1 gene were generated under the control of a promoter from the maize ubiquitin gene. Three homozygous lines of $T_2$ progenies with single transgene insert were chosen for gene expression analysis. RT-PCR and western blot analysis results indicated that the OsBADH1 gene was effectively expressed in transgenic tobacco leading to the accumulation of glycine betaine. Transgenic lines demonstrated normal seed germination and morphology, and normal growth rates of seedlings under salt stress conditions. These results suggest that the OsBADH1 gene could be an excellent candidate for producing plants with osmotic stress tolerance.