• Title/Summary/Keyword: transgenic

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Preselection and cloning of transgenic emb (유전자전환 수정란의 선별과 복제)

  • Lee, Hyo-Jong
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 1998.05a
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    • pp.12-28
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    • 1998
  • The technology of creating transgenic animals has a potential value in improving productivity and disease resistance of animals, gene therapy, drug pharming and production of model animals for certain diseases. Up to date, fairly low success rate of production of transgenic animals and a pronounced variability with respect to the expression of transgenes have been much observed. The mechanisms how to integrate the injected genes with a certain part of the genomes are unknown yet. Many techniques in gene transfer, beside microinjection, have been introduced and explored thus to improve the production efficiency of transgenic animals. In this article, the methods and efficiency of gene-transfer techniques, the detection and preselection of transgenes in embryos by PCR- and GFP-screenings and cloning of preselected transgenic embryos by nuclear transplantation are described and discussed. Some experimental results showed that the early screening and selection of integration of the injected gene with embryonic genome by polymerase chain reaction(PCR) and green fluorecence protein(GFP) were promising methods. Further, the application of nuclear transplantation technology to cloning and multiplication of the positively integrated genes in the cleaving embryos and embryonic cells will be beneficially used for the mass production of transgenic embryos and consequently improving the production efficiency in transgenic animals.

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Phenotype Comparison between Herbicide Tolerant Transgenic Rice and Weedy Rice (제초제 내성 형질전환 벼와 잡초성 벼의 표현형질 비교)

  • Go, Eun Mi;An, Joo Hee;Nam, Ki Jung;Nam, Kyong-Hee;Park, Kee Woong;Back, Kyoungwhan;Kim, Chang-Gi
    • Weed & Turfgrass Science
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    • v.2 no.1
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    • pp.15-22
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    • 2013
  • The phenotypic traits of herbicide-tolerant transgenic rice were compared with those of wild type (Dongjin) as well as two accessions (Hwaseong-aengmi 1 and Gwangyang-aengmi 12) of weedy rice. This study was conducted to investigate whether unintentional alterations in phenotypic characteristics occurred in the transgenic rice and whether the altered traits were similar to those in the two weedy rices. All qualitative traits studied were similar in the transgenic or wild-type rice. On the other hand, awn presence, flag leaf attitude and grain color differed considerably between herbicide-tolerant transgenic rice and weedy rice. As for quantitative traits, plant height, the number of tillers per plant and shoot dry weight were significantly greater for weedy rice than transgenic or wild-type rice. Grain weight per plant and 1000-grain weight of transgenic (or wild-type) rice were significantly greater than those of weedy rice. Transgenic rice shattered less than the other rices. Amylose and protein contents in embryos of transgenic rice were significantly different from those of weedy rice. The potential for weediness of the transgenic rice may be assessed using phenotype comparison between transgenic and weedy rice as shown in this study.

Expression Analysis of Diphtheria Toxin-A Gene Regulated by Lck Promoter in Transgenic Mice (형질전환생쥐에서 Lck Promoter에 의한 Diphtheria Toxin-A Gene의 발현 분석)

  • 나루세겐지;이승현;최화식;이성호;박창식;진동일
    • Korean Journal of Animal Reproduction
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    • v.27 no.3
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    • pp.225-231
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    • 2003
  • Transgenic mice containing Diphtheria Toxin-A (DT-A) gene fused to proximal lck promoter sequences was used for analysis of DT-A gene expression and thymocyte development. The diphtheria toxin gene was expressed in thymus, spleen and liver of transgenic mice confirmed by RT-PCR and Northern blotting. A FACS analysis with thymocyte cell surface antigens antibodies (CD4 and CD8) showed that the number of peripheral mature single positive thymocytes ($CD4^{+}\;and\;CD8^{+}$ cells) T-cells was severely reduced in transgenic mice compared to that in the non-transgenic littermates. A relative portion of $CD8^{+}$ single positive thymocytes was about 33.2% in transgenic peripheral T-cells while 50.6% in wild type. Reduction of $CD4^{+}$ cell numbers in transgenic mice was observed (5.9% in transgenic versus 10.3% in non-transgenic). The data from analysis of these transgenic mice indicate that the proximal lck promoter regulated the expression of DT-A gene at high level in developing thymocytes and the DT-A disrupted developing thymocytes in transgenic mice.

Field Performance and Morphological Characterization of Transgenic Codonopsis lanceolata Expressing $\gamma-TMT$ Gene.

  • Ghimire, Bimal Kumar;Li, Cheng Hao;Kil, Hyun-Young;Kim, Na-Young;Lim, Jung-Dae;Kim, Jae-Kwang;Kim, Myong-Jo;Chung, Ill-Min;Lee, Sun-Joo;Eom, Seok-Hyun;Cho, Dong-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.5
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    • pp.339-345
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    • 2007
  • Field performance and morphological characterization was conducted on seven transgenic lines of Codonopsis lanceolata expressing ${\gamma}-TMT$ gene. The shoots were obtained from leaf explants after co-cultivation with Agrobacterium tume-faciens strain LBA 4404 harboring a binary vector pYBI 121 that carried genes encoding ${\gamma}-Tocopherol$ methyltransferase gene (${\gamma}-TMT$) and a neomycin phosphotransferase II gene (npt II) for kanamycin resistance. The transgenic plants were transferred to a green house for acclimation. Integration of T-DNA into the $T_0\;and\;T_1$ generation of transgenic Codonopsis lanceolata genome was confirmed by the polymerase chain reaction and southern blot analysis. The progenies of transgenic plants showed phenotypic differences within the different lines and with relative to control plants. When grown in field, the transgenic plants in general exhibited increased fertility, significant improvement in the shoot weight, root weight, shoot height and rachis length with relation to the control plants. However, all seven independently derived transgenic lines produced normal flower with respect to its shape, size, color and seeds number at its maturity. Indicating that the addition of a selectable marker gene in the plant genome does not effect on seed germination and agronomic performance of transgenic Codonopsis lanceolata. $T_1$ progenies of these plants were obtained and evaluated together with control plant in a field experiment. Overall, the agronomic performance of $T_1$ progenies of transgenic Codonopsis lanceolata showed superior to that of the seed derived non-transgenic plant. In this study, we report on the morphological variation and agronomic performance of transgenic Codonopsis lanceolata developed by Agrobacterium transformation.

Effect of Feeding Transgenic Cottonseed vis-à-vis Non-transgenic Cottonseed on Haematobiochemical Constituents in Lactating Murrah Buffaloes

  • Singh, Maha;Tiwari, D.P.;Kumar, Anil;Kumar, M. Ravi
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.12
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    • pp.1732-1737
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    • 2003
  • An experiment was conducted to investigate the effect of feeding transgenic cottonseed (Bt.) vis-a-vis non-transgenic (non-Bt.) cottonseed on blood biochemical constituents in lactating Murrah buffaloes. Twenty Murrah buffaloes in mid-lactation were divided into 2 groups of 10 each. Animals of group I were fed with 39.5% non-transgenic cottonseed in concentrate mixture while the same percentage of transgenic (Bt.) cottonseed was included in the concentrate mixture fed to the animals of group II. Animals of both groups were fed with concentrate mixture to support their milk production requirements. Each buffalo was also offered 20 kg mixed green fodder (oats and berseem) and wheat straw ad libitum. The experimental feeding trial lasted for 35 days. There was no significant difference in the dry matter intake between the two groups of buffaloes. All the buffaloes gained body weight, however, the differences were non significant. Total erythrocyte count, hemoglobin content and packed cell volume were $9.27{\pm}0.70${\times}10^6/{\mu}l$, $13.01{\pm}0.60gdl$ and $34.87{\pm}1.47%$, respectively in group I with the corresponding figures of $8.88{\pm}0.33$, $12.99{\pm}0.52$ and $31.08{\pm}1.52$ in group II. The values of total erythrocyte count, haemoglobin content and packed cell volume did not differ significantly between the two groups of buffaloes. The concentration of plasma glucose, serum total proteins, albumin, globulin, triglycerides and high density lipoprotein were non significantly higher in buffaloes fed non-transgenic cottonseed than in buffaloes fed transgenic cottonseed. The cholesterol concentration was significantly (p<0.01) higher in buffaloes of group I ($136.84{\pm}8.40mg/dl$) than in buffaloes of group II ($105.20{\pm}1.85mg/dl$). The serum alkaline phosphotase, glutamic-oxaloacetate transaminase and glutamic-pyruate transaminase activities did not differ significantly between two groups of buffaloes. However, serum glutamic-pyruate transaminase activity was considerably high in buffaloes fed nontransgenic cottonseed as compared to buffaloes fed transgenic cottonseed. Bt. proteins in serum samples of animals of group II were not detected after 35 days of feeding trial. It was concluded that transgenic cottonseed and non-transgenic cottonseed have similar nutritional value without any adverse effects on health status of buffaloes as assessed from haematobiochemical constituents.

Activation by Combined Treatment with Cycloheximide and Electrical Stimulation of In-Vitro Matured Porcine Oocytes Improves Subsequent Parthenogenetic Development

  • Naruse Kenji;Kim Hong-Rye;Shin Young-Min;Chang Suk-Min;Lee Hye-Ran;Tarte Vaishali;Quan Yan-Shi;Kim Beak-Chul;Park Tae-Young;Choi Su-Min;Park Chang-Sik;Jin Dong-Il
    • Reproductive and Developmental Biology
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    • v.30 no.1
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    • pp.41-45
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    • 2006
  • Electrical treatment has been widely used for porcine oocytes activation. However, developmental rates following electrical activation of porcine oocytes is relatively inefficient compared to other domestic animals. To investigate the effects of porcine oocytes on combined activation by both chemical and electrical treatment, in-vitro matured oocytes were activated by combined cycloheximide and electrical pulses treatment. Cumulus-free oocytes were exposed with NCSU-23 medium containing cycloheximide $(10{\mu}g/ml)$ for 0, 5, 10, 20, 30 min and then activated by electrical pulse treatment and cultured in PZM-3 for 8 days. Also effects of exposure to $6.25{\mu}M$ calcium ionophore for 2 min for cumulus-free oocytes were tested. The percentage of blastocyst formation in 10 min exposure to $10{\mu}g/ml$ cycloheximide and electrical pulse treatment was significantly increased (P<0.05) than in the control group. And exposure to $6.25{\mu}M$ calcium ionophore for 2 min with $10{\mu}g/ml$ cycloheximide for 10min and electrical pulse treatment significantly increased (P<0.05) the percentage of blastocyst developmental rates than the control group. In conclusion, activation by combined cycloheximide and electrical stimulation treatment promoted the subsequent development of porcine oocytes and improved the subsequence blastocyst development.

Approaches to Improving Production Efficiencies of Transgenic Animals

  • Tojo, Hideaki
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.343-346
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    • 2000
  • Transgenic animals are very useful for scientific, pharmaceutical, and agricultural purposes. In livestock, transgenic technology has been used forth genetic alteration of farm animals, the production of human proteins inlarge quantities in the milk of transgenic farm animals (Clark et al., 1989; Ebert et al., 1991; Kimpenfort et al., 1991; Wall et al., 1991; Kimpenfort et al., Well et al,m 1991; Hill et al., 1992; Velander et al., 1994; Chen et al.), and the generation of animals with organs suitable for xenotransplantation (Pinkert, 1994; Chen et al., 1999). (omitted)

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