• 한국수정란이식학회지
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• 제14권1호
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• pp.1-8
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• 1999

The efficiency of transgenic livestock animal production may be improved by early selection of transgenci preimplantation embryos. To examine the possibility of GFP gene as a non-invasive marker for the early screening of transgenic embryo, the GFP gene was microinjected into rabbit zygotes and the later stages of preimplantation embryos were examined for the expression of GFP. The presence of injected DNA was detected by PCR analysis and the expression of GFP was detected by observing green fluorescence in embryos under a fluorescent microscope. Out of 108 GFP gene-injected rabbit zygotes, seventy three(67.6%) were fluorescence-positive. When 11 fluroresecence-positive blastocysts were analyzed for the presence of GFP gene by PCR, 6(54.5%) were positive, and all of the 8 flrouescence-negative blastocysts were also negative by PCR. The results indicate that the screening of transgene in rabbit embryos by PCR analysis and GFP detection could be a promising method for the preselection of transgenic embryos.

• Reproductive and Developmental Biology
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• 제40권1호
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• pp.7-13
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• 2016

Effectiveness of transgene transfer into genome is crucially concerned in mass production of the bio-pharmaceuticals using genetically modified transgenic animals as a bioreactor. Recently, the mammary gland has been considered as a potential bioreactor for the mass production of the bio-pharmaceuticals, which appears to be capable of appropriate post-translational modifications of recombinant proteins. The mammary gland tissue specific vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals. In this study, to minimize physiological disturbance caused by constitutive over-expression of the exogenous gene, we constructed new retrovirus vector system designed for mammary gland-specific expression of the hEPO gene. Using piggyBac vector system, we designed to express hEPO gene under the control of mammary gland tissue specific and lactogenic hormonal inducible goat ${\beta}$-casein or mouse Whey Acidic Protein (mWAP) promoter. Inducible expression of the hEPO gene was confirmed using RT-PCR and ELISA in the mouse mammary gland cells treated with lactogenic hormone. We expect the vector system may optimize production efficiency of transgenic animal and reduce the risk of global expression of transgene.

• Journal of Plant Biotechnology
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• 제5권1호
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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.

• Journal of Plant Biotechnology
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• 제39권4호
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• pp.242-252
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• 2012

Seed storage oils are essential resources for not only human and animal diets but also industrial applications. The primary goal of this study was to increase seed oil content through comparative analysis of two seed-specific promoters, AtFAE1 from Arabidopsis Fatty Acid Elongase 1 gene and BnNapin from Brassica napus seed storage protein gene. AtWRI1 and AtDGAT1 genes encoding an AP2-type transcription factor and a Diacylglycerol Acyltransferase 1 enzyme, respectively, were expressed under the control of AtFAE1 and BnNapin promoters in Arabidopsis. The total seed oil content in all transgenic plants was increased by 8-11% compared with wild-type seeds. The increased level of oil content in AtWRI1 and AtDGAT1 transgenic lines under the control of both promoters was similar, although the activity of the BnNapin promoter is much stronger than that of AtFAE1 promoter in the mature stage of developing seeds where storage oil biosynthesis occurs at a maximum rate. This result demonstrates that the AtFAE1 promoter as well as the BnNapin promoter can be used to increase the seed oil content in transgenic plants.

• Journal of Plant Biotechnology
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• 제34권1호
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• pp.11-17
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• 2007

수박작물의 대목용으로 사용하는 수박공대에 CGiMMV-CP 유전자를 도입하여 개발된 LM 수박공대의 CGMMV 내성 정도를 격리하우스와 노지 포장내에서 조사하였다. 격리온실에서의 $T_{3}$ 형질전환 수박대목의 CGMMV 내성은 접종 후 70일까지 유지되는 반면 대조구는 접종 후 20일에 전부 이병되었다. 인위적 토양전염 포장에서 형질전환체는 접종후71일까지 약 40%의 내성률을 보였으며, 대조구는 접종 후 37일에 모두 이병되었다. 인위적 접촉전염 포장에서 형질전환체는 대조구에 비해 약 10일 정도 지연효과를 보였다. 따라서 CGMMV-CP 형질전환체는 CGMMV에 저항성을 가진 것이 아니라 감염시기를 지연시키는 부분 내성으로 나타났다. CGMMV-CP homozygous T 세대를 진전시켜서 형질전환 수박공대 계통을 $BC_{1}T_{5}$ 세대에서 선발하였다. 또한 LM 수박공대에 형질전환 되지 않은 접수 (슈퍼금천수박)를 접목하여 non-LM 수박을 생산하고 CGMMV-CP 유전자에 관련된 물질의 이동 여부를 조사하였다. PCR, northern, western 분석한 결과 수박공대 대목에서 형성되는 DNA, RNA, protein 물질이 접수로 이동되지 않음을 확인하였다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.165-165
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• 2017

Abiotic stress is one of the major serious limiting factors in rice (Oryza sativa) and caused rice production losses. It is important to precisely screen valuable genetic resources for improving stress tolerance and understanding tolerance mechanism to abiotic stresses. Because there are differences of experiment designs for screening of tolerant plant in several studies related to abiotic stress, this study has performed to provide the rapid and efficiency screening method for selection of tolerance rice to drought and salinity stresses. Two week-old rice seedlings that reached about three leaf stage were treated with drought and salinity stresses and examined tolerant levels with tolerant and susceptible control varieties, and transgenic plants. To determine the optimum concentration for the selection of drought and salinity condition, tolerant, susceptible and wild-type plants were grown under three soil moisture contents (5, 10 and 20% water contents) and three NaCl concentrations (100, 200 and 250 mM) for 10 days at seedling stage. 200 mM NaCl concentration and 5% moisture content soil were determined as the optimum conditions, respectively. The described methodologies in this study are simple and efficiency and might help the selection of drought and salinity tolerance plants at the 3,4-leaf-seedling stage.

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2003년도 추계학술대회
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• pp.109-110
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• 2003

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.213-213
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• 2004

Exogeneous DNA can reproducibly be delivered by co-injected spermatozoa and this transgenesis method is very efficient protocol. But, mosaic patterns of transgenic embryos and offspring were shown frequently. Whole blastomere integration is important in transgenic animal economics. (omitted)

• 한국가금학회지
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• 제26권2호
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• pp.109-118
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• 1999

Hen's eggs have been regarded as one of the best animal bioreactors to produce biologically active peptides originated from many organisms including human. Despite the last decade's efforts to produce transgenic chicken for any commercial purposes, the results so far reported are very disappointing, indicating that hen's eggs are very difficult to crack for transgenesis. Comparatively large female gamete with enormous amount of yolk may be one of the major obstacles in achieving a similar feat to those of other vertebrate species including mouse, sheep, fish and frog. The delay or less efficiency evidenced may instruct to try an alternative way of gens transfer into chicken egg. Sperm-mediated gene transfer is one of them, and may require a great deal of understanding of mechanisms involved in early fertilization and embryonic development. In other animals where the technique was successful, basic mechanisms have been well studied and established only by painstaking efforts for decades. This paper discusses the accumulated knowledge on early fertilization mechanism in the chicken and how can this information be utilitzed to find the alternative gene transfer in making transgenic chicken.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.94-94
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• 2002

Increasing competition in the livestock industry has forced producers to cut costs by adopting new technologies aimed out increasing production efficiency. Non-ruminant livestock do not express fibrolytic enzymes. The major plant cell wall components of cereals, primarily β-glucans and arabinoxylans, form gel-like structures in the small intestines that trap nutrients. The viscous polysaccharides can also cause severe gastrointestinal disorders. (omitted)