• Title/Summary/Keyword: Transgenic line

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Difference in Physiological Responses to Environmental Stress in Protox Inhibitor Herbicide-Resistant Transgenic Rice and Non-transgenic Rice (Protox 저해형 제초제 저항성 형질전환벼와 비형질전환벼의 환경스트레스에 대한 생리적 반응 차이)

  • Yun, Young-Beom;Kwon, Oh-Do;Shin, Dong-Young;Hyun, Kyu-Hwan;Lee, Do-Jin;Jung, Ha-Il;Kuk, Yong-In
    • Korean Journal of Weed Science
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    • v.32 no.1
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    • pp.35-43
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    • 2012
  • The objective of this research was to confirm the difference in physiological responses to environmental stresses such as chilling, high temperature, NaCl, and chemical stress (paraquat) in Protox inhibitor resistant-transgenic rice (MX, PX, and AP37) and its non-transgenic counterpart (WT). Transgenic and non-transgenic rice plants were exposed to a chilling temperature of $5^{\circ}C$ for 1 day or a high temperature of $45^{\circ}C$ for 4 days and allowed to recover at $25^{\circ}C$ for 6 days after the chilling treatment or 8 days after the high temperature treatment. Leaf injury, shoot fresh weight, porphyrin biosynthesis substances, and chlorophyll content were investigated in transgenic and non-transgenic rice at 6 days after 0.5% and 1% NaCl treatments or at 5 days after 0~300 ${\mu}M$ paraquat treatments. No significant difference in leaf injury and shoot fresh weight were observed between transgenic and non-transgenic rice during chilling and recovery. Plant height and shoot fresh weight were also similar between transgenic and non-transgenic rice during the high temperature and recovery period (0~5 days). However, plant height and shoot fresh weight in transgenic rice line MX and PX were lower than in non-transgenic rice at 6 days for recovery. Leaf injury, chlorophyll, and Mg-Proto IX ME contents had no significant difference between transgenic rice and non-transgenic rice after NaCl treatment, but Proto IX content for AP37 and shoot fresh weight for PX and AP37 in 0.5% NaCl treatment were significantly reduced compared with non-transgenic rice. There was no difference in leaf injury and shoot fresh weight when comparing transgenic rice and non-transgenic rice after paraquat treatment. Although transgenic rice and non-transgenic rice showed a little difference at a particular measurement period in certain environmental stresses, there was generally no difference in physiological responses between transgenic rice and non-transgenic rice.

Seed longevity of glyphosate resistant transgenic creeping bentgrass (Agrostis stolonifera L.) lines

  • Hancock, Daniel;Park, Kee Woong;Mallory-Smith, Carol A.
    • Journal of Ecology and Environment
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    • v.38 no.4
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    • pp.437-442
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    • 2015
  • Studies to estimate seed longevity and dormancy of creeping bentgrass (Agrostis stolonifera L.) were conducted from 2000 to 2005 at Corvallis and Hermiston, Oregon. Seeds from three transgenic glyphosate resistant creeping bentgrass lines, 48-10, 48-13, and ASR368, and one non-transgenic glyphosate susceptible line, SR1020, were used. Creeping bentgrass seeds were buried at 3, 18 and 31 cm in 2000 and removed 6, 12, 18, 24, and 51 months later. Soil type and climatic conditions were different at the two locations. At Corvallis, the soil was a Malabon silty clay loam, and the winters wet and mild. The soil at Hermiston was an Adkins fine sandy loam, and winters drier and colder. Seeds of all creeping bentgrass lines deteriorated faster at Corvallis than at Hermiston. The estimated half-lives of creeping bentgrass lines buried at Corvallis were 8.4 to 20.2 months, while those buried at Hermiston were 8.4 to 37.7 months. At both sites, seeds of the glyphosate resistant lines, 48-10 and 48-13, deteriorated faster than the susceptible line, SR1020. However, seed deterioration in the resistant line, ASR368, was slower than all other creeping bentgrass lines. Based on the germination test, exhumed intact seeds at Corvallis were more dormant than those at Hermiston. If buried, it could be expected that viable creeping bentgrass seeds will persist more than 4 years after the seeds are introduced to a site, but environmental conditions can influence both seed longevity and dormancy.

Transposon piggyBac mediated Ipax6 Expression in Malaria Vector Anopheles stephensi (말라리아 매개 모기 Anopheles stephensi에서 트랜스포존 piggyBac을 이용한 Pax6 발현)

  • Koo Hyeyoung
    • Development and Reproduction
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    • v.8 no.1
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    • pp.19-25
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    • 2004
  • Pax6, a member of the highly conserved homeobox gene family, is known to be expressed in spatially and temporally restricted pattern during embryogenesis. To examine the spatial expression pattern of Pax6 in malaria vector mosquito Anopheles stephemi, in different molecular environment, the germ line transformation technique using piggyBac transposon combined with the use of Pax6 specific 3xp3-EGFP marker was utilized. Four transgenic lines with a transformation rate of 6.7% were established. Transgenes were stably expressed in subsequent several generations. The transgenic lines showed 3 different expression pattern with spatial specificity, possibly due to enhancing and/or silencing position effects. In two transgenic lines, noble expression pattern of Pax6 was observed in the region that has not been previously reported in any animal species. The results show that the tranposon piggyBac mediated germ line transformation system can be used as an efficient tool for the generation of diverse spatially restricted reporter gene expression.

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Germ-line Transmission of Pseudotyped Retroviral Vector in Chicken

  • Heo, Y.T.;Kim, T.;Lee, Y.M.;Lee, C.K.;Kwon, M.S.;Koo, B.C.;Roh, K.S.;Whang, K.;Han, D.W.;Chung, K.S.;Lee, H.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.1
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    • pp.27-32
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    • 2004
  • Using MLV (murine leukemia virus)-based retrovirus vectors encapsidated with VSV-G (vesicular stomatitis virus G glycoprotein), we tried to make transgenic chickens carrying the transferred genes in their chromosomes. Twenty one days after virus injection beneath the blastoderms of unincubated chicken embryos (stage Ⅹ, at laying), DNA isolated from the hatched chicks were analyzed by PCR with two sets of primers specific for EGFP (enhanced green fluorescence protein) gene or $Neo^R$ (E. coli neomycin resistant) gene. Among sixty-seven embryos injected with retrovirus, four of them were identified to carry the EGFP genes in their genomes. Remarkably, one transgenic chick showed presence of the retrovirus vector sequences in all organs differentiated from one of endoderm, mesoderm, and ectoderm. Expression of EGFP gene was not detected, however, the stable germ line transmission of transgene was verified in spermatozoa from the founder chicken and 50% of $F_1$ progenies.

The increased GUS gene inactivation over generation in Arabidopsis transgenic lines (애기장대 형질전환 식물체의 세대경과에 따른 GUS유전자의 비활성화에 관한 연구)

  • Park, Soon-Ki
    • Journal of Life Science
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    • v.12 no.1
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    • pp.67-76
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    • 2002
  • The effect of transgene inactivation in T2, T3 and F2 generations was analyzed in progeny seedlings which had been generated by Agrobacterium (LBA4404/pBI121)-mediated transformation in Arabidopsis thaliana. In a system which investigated in the expression of $\beta$-glucuronidase(GUS)gene in kanamycin-resistant (ke $n^{R}$)seedlings, GUS inactivated seedlings were observed in 5 of 12 tested lines of T2 generation and the frequency of GUS inactivation was approximately 2.3%. Lines with multi-copies of T-DNA exhibited severe GUS gene inactivation with the frequency of 5.8% in T2 generation. In T3 generation lines exhibited GUS gene inactivation with the frequency of 1.3%. In contrast, inactivation increased dramatically up to 12.6% in multi-copy T-DNA line. A similar phenomenon was also found in F2 progeny from a transgenic line which had been crossed with wild-type Arabidopsis plant, WS-O (GUS gene inactivation frequency 9.9%). These results indicate that the foreign gene introduced into the plant was inactivated progressively in its transmission during subsequent generations and the transgenic line with multi-copies of T-DNA tended to show more increased inactivation.

바이러스 외피단백질 유전자로 형질전환된 연초 식물체의 TMV 저항성 발현 및 유전자 안정성

  • 박성원;이기원;이청호;이영기;강신웅;최순용
    • Journal of the Korean Society of Tobacco Science
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    • v.21 no.1
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    • pp.77-81
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    • 1999
  • Tobacco plants(Nicotiana tabacum cv. NC82) transformed with TMV CP cDNA were self-fertilized until 8th generation (R$_{8}$), and the transgenic plants from 6th to 8th generation were analized for their resistance to tobacco mosaic virus(TMV) and stability of the gene expression. The 6th generation of the plants(R$_{6}$) showed high resistance(81-91 %) to TMV at eight weeks after artificial inoculation with the virus. The transgenic cell line 601 was the most prominant in the expression of resistance. 98 % of the plants showed no symptom without any agronomic phynotepe variation when they were inoculated with the virus in a experimental field. However, 2% of the plants were revealed as delay type of symptom with mild mosaic on a few leaves. The viral resistance in greenhouse tests of the 7th generation (R$_{7}$) was 54-64%, and the number of delay type plants were increased than that of 6th generation plants. In the 8th generation, 81 % of the plants was complete resistant to the virus. The TMV CP cDNA of the transgenic plants of each generation was also confirmed by genomic PCR, and there was no systemic viral multiplication in the resistant plants. It suggests that the viral resistance and gene expression of the transgenic plants might be stable through the generations.ons.s.

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Formation of Brain Tumor and Lymphoma by Deregulation of Apoptosis Related Gene Expression in VP-SV40 T Ag Transgenic Mice

  • Lee, Jeong-Woong;Lee, Eun-Ju;Lee, Hoon-Taek;Chung, Kil-Saeng;Ryoo, Zae-Yoong
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.47-47
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    • 2001
  • The neuropeptide vasopressin (VP) is a nine- amino acid hormone synthesized as preprohormone in the cell bodies of hypothalamic magnocellular neurons. The tumor in magnocellular neurons of the hypothalamus is associated with disfunctions of the cell bodies, leading to the diabetes insipidus. In order to study with the diabetes insipidus caused by a defect in VP synthesis and its secretion, we have produced the transgenic mice regulated by vasopressin promoter inserted to SV40 T antigen coding sequence (pVPSV.IGR2.1). One transgenic line expressing high levels of SV40 T antigen was propagated. The founder and all transgene positive adult animals have appeared with shorten mortality or apparent phenotypic abnormalities, including immune complex disease, and eventually die between 4 and 8 months of age. The mRNA and protein of SV40T antigen transgene were detected in brain of fetus as well as in brain, spleen, lung and lymph node in moribund at the age of 20 weeks. Histological analysis of transgenic mice showed that tumor developed in brain similar to primitive neuroectodermal tumors (PNET) in man. We also detected lymphomas in spleen and lymph node, and consequent tumor formation in various tissues of the transgenic mice. In pVPSV.IGR2.1, 21% mice showed brain tumor (PNET) at 5 weeks and 100% mice showed brain tumor after 15 weeks. In addition, Expression of apoptosis related genes (Bcl-28 & Bax) was increased over their age in mice with PNET as compared to control mice. Apoptosis related gene expression might be deregulated in mice with brain tumor. However, transgenic mice were not developed with the diabetes insipidus. These mice represent the first disease model to exhibit primitive neuroectodermal tumor in brain, as well as a unique model system for exploring the cellular pathogenesis of lymphomas.

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Fungal pathogen protection in transgenic lettuce by expression of a apoptosis related Bcl-2 gene (Apoptosis 관련 Bcl-2유전자의 도입을 통한 곰팡이 저항성 형질전환 상추의 육성)

  • Seo, Kyung-Sun;Min, Byung-Whan
    • Journal of Plant Biotechnology
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    • v.38 no.3
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    • pp.209-214
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    • 2011
  • Transgenic lettuce plants were successfully obtained from hypocotyl explants inoculated with Agrobacterium tumefaciens, which harbored a binary vector plasmid with Bcl-2 gene, related to apoptosis. After culture and selection on MS medium a number of kanamycin-resistant plantlets were regenerated. Polymerase chain reaction, Southern blot analysis and Northern blot analysis were used to identify and characterize the transgenic plants with the integrated Bcl-2 gene. Over 100 transgenic plants have been established in soil and flowered in the greenhouse. T1 progeny of 100 transgenic lettuce inbred lines were inoculated with Sclerotinia sclerotiorum. Expression of the Bcl-2 peptide in transgenic lettuce plants provides high levels of field resistance against Sclerotinia sclerotiorum, causal agent of the agronomically important fungal disease of lettuce.

Stable Transmission and Expression of TPO Transgene up to 10 Generation in the Transgenic Mice (형질전환 생쥐에서 제10세대까지 TPO 유전자의 안정적 전이와 지속적인 발현)

  • 정진우;오건봉;한용만;이경광
    • Korean Journal of Animal Reproduction
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    • v.27 no.1
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    • pp.9-14
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    • 2003
  • The pBT-L transgenic mice carrying human TPO gene in conjunction with bovine $\beta$-casein promoter express human TPO in milk during lactation. In this study, stability of germ line transmission and expression of pBT-L transgene integrated into host chromosome were monitored up to generation F10 of transgenic pBT-L/15 line. When male mouse of generation F8 was crossbred with normal females, approximately half of offsprings (51.3$\pm$18.98%) were identified as transgenic mice. Generation F9 and F 10 mice also showed similar transmission rates (43.8$\pm$18.98% and 71.4$\pm$26.98%, respectively), implying that pBT-L transgene can be transmitted stably up to long term generation in the transgenic mice. Expression levels of human TPO from milk of generation F9 and F10 mice were 1.1$\pm$0.33 mg/ml and 1.1 $\pm$0.45 mg/ml, respectively, which are similar to expression level of generation F2 mice. In conclusion, our results suggest that transgenic animals once established will continuously pass their transgenes to the progeny through the breeding program with the same productivity of human protein in their milk.

Proteomic Analysis and the Antimetastatic Effect of N-(4methyl)phenyl-O-(4-methoxy) phenyl-thionocarbamate-Induced Apoptosis in Human Melanoma SK-MEL-28 cells

  • Choi Su-La;Choi Yun-Sil;Kim Young-Kwan;Sung Nack-Do;Kho Chang-Won;Park Byong-Chul;Kim Eun-Mi;Lee Jung-Hyung;Kim Kyung-Mee;Kim Min-Yung;Myung Pyung-Keun
    • Archives of Pharmacal Research
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    • v.29 no.3
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    • pp.224-234
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    • 2006
  • We employed human SK-MEL-28 cells as a model system to identify cellular proteins that accompany N-(4-methyl)phenyl-O-(4-methoxy)phenyl-thionocarbamate (MMTC)-induced apoptosis based on a proteomic approach. Cell viability tests revealed that SK-MEL-28 skin cancer cells underwent more cell death than normal HaCaT cells in a dose-dependent manner after treatment with MMTC. Two-dimensional electrophoresis in conjunction with matrixassisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry analysis or computer matching with a protein database further revealed that the MMTC-induced apoptosis is accompanied by increased levels of caspase-1, checkpoint suppressor-1, caspase-4, NF-kB inhibitor, AP-2, c-Jun-N-terminal kinase, melanoma inhibitor, granzyme K, G1/S specific cyclin D3, cystein rich protein, Ras-related protein Rab-37 or Ras-related protein Rab-13, and reduced levels of EMS (oncogene), ATP synthase, tyrosine-phosphatase, Cdc25c, 14-3-3 protein or specific structure of nuclear receptor. The migration suppressing effect of MMTC on SK-MEL-28 cell was tested. MMTC suppressed the metastasis of SK-MEL-8 cells. It was also identified that MMTC had little angiogenic effect because it did not suppress the proliferation of HUVEC cell line. These results suggest that MMTC is a novel chemotherapeutic and metastatic agents against the SK-MEL-28 human melanoma cell line.