• Title/Summary/Keyword: 형질 세포

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Effects of Gamma-Irradiation on the Sterilization of Primordial Germ Cells in Quail (메추리 원시생식세포 감소를 위한 감마선 조사의 효과)

  • Park, Kyung-Je;Kim, Tae-Min;Lee, Hyung-Chul;Jang, Hyun-Jun;Song, Gwon-Hwa;Han, Jae-Yong
    • Korean Journal of Poultry Science
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    • v.37 no.2
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    • pp.139-143
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    • 2010
  • Quail is a very useful animal model for studying vertebrate development because of its small body size and unique reproductive traits. This species is also ideal model for producing germline chimeras via transferring exogenous primordial germ cells (PGCs) into the recipient embryo. To increase the contribution efficiency of donor PGCs into recipients' tissues, decreasing the population of endogenous PGCs has been rate-limiting factor. We therefore conducted this study to investigate if gamma ($\gamma$)-irradiation depletes endogenous PGCs in developing quail embryo. Firstly, freshly laid stage X quail embryos were irradiated with various output of $\gamma$-irradiation and its teratogenic effect on the embryo was evaluated. Although a dose-dependent increase in the number of embryo showing malformation was found as the output increased (0, 250, 500, 750, and 1,000 rads), only a maximum of 10.1% of embryos were abnormal in 1,000 rads. Immunocytochemical analysis using the QCR1 antibody, which is specific marker for quail PGCs, was conducted to analyze the effect of sterilization. As results, $\gamma$-rays at a dose-rate of 500 rads/73 sec onto undeveloped stage X embryo significantly reduced the number of germ cells to an average of 75.55 % and 82.03 % in male and female embryos, respectively. We conclude that $\gamma$-ray selectively targets PGCs while affects minimally to the somatic development in quail embryo. Our results will not only provide important data for germline chimera production but can be used for analyzing the effect of ionized rays on the differentiating germ cells in various stages during animal development.

Targeted Gene Knockout in Chickens Mediated by TALENs (연구 - 유전자 가위 기법을 활용한 오브알부민 생산 유전자를 제거한 유전자 변형 닭 생산)

  • Han, Jae-Yong;Park, Tae-Seop
    • KOREAN POULTRY JOURNAL
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    • v.46 no.10
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    • pp.132-133
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    • 2014
  • 생식선 줄기세포 원천기술을 이용한 형질전환 조류 생산 시스템과 고효율의 유전자적 중 기술인 유전자가위법(TALEN)을 도입함으로써 기초연구 및 단기간내 새로운 가금품종 개발이 가능하게 되었다. 또한, 신약개발 및 치료물질 대량생산을 위한 형질전환가금품종 개발에 응용될 수도 있어 축산과 더불어 의약, 약학 등 매우 다양한 분야에서 가금의 활용 범위를 넓힐 수 있을것으로 기대된다. 개발된 기술은 계란성분 조절을 통한 기능성 식품 및 단백질-신약을 포함한 신물질 생산을 목적으로 하는 지식기반 생명산업의 획기적인 발전을 유도할 수 있다. 이에 유전자 가위 기법을 활용한 오브알부민 생산 유전자를 제거한 유전자 변형 닭 생산에 관한 주요내용을 소개코자 한다.

Expression of GFP in Various Cells and Tissues Isolated from Recloned GEP-Transgenic Pigs

  • Park, K.W.;Lee, H.;Kim, S.U.;Choi, S.S.;Choi, S.K.;Chung, Y.S.;Hong, J.Y.;Hong, S.;Jung, J.Y.;Park, C.S.;Lee, S.H.
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.188-188
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    • 2004
  • 근래에 많은 형질전환 실험동물이 생산되어 인간을 대체하여 전임상실험에 사용되고 있으며, 그 요구는 점차 증가되고 있다. 그러나 지금까지 생산된 것은 소형 실험동물에 국한되고 있어, 사람과의 그 크기와 기능에 거리가 있었던 것이 사실이다. 본 실험은 중형 실험동물이자 사람과 여러 기관 및 조직이 유사하다고 알려진 돼지로부터 난자를 채취하여 난자의 핵을 GFP가 발현된 세포의 핵으로 치환하여 태어난 산자를 재복제하여 생산된 돼지 신생자를 이용하여 각 기관 및 조직에서 GFP의 발현을 RT-PCR, Northern blot 및 immunocytochemistry 방법으로 분석하였다. (중략)

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Gonadotropins : Basic View and Gene Expression (성선자극호르몬 : 유전자 발현에 대한 고찰)

  • Yukio Kato;Koichiro Gen;;Takako Kato
    • Korean Journal of Animal Reproduction
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    • v.19 no.1
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    • pp.15-34
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    • 1995
  • 1970년말부터 뇌하수체 성선자극호르몬(gonadotropic hormone ; GTH)의 유전자 구조(FSH$\beta$, LH$\beta$ 및 공동의 $\alpha$쇄)가 다양한 종에서 밝혀지기 시작하였으나 이러한 유전자의 조직/세포 특이적 분비양식과 세포외 신호에 의한 조절양식은 정확히 밝혀져 있지 않다. 그러나 최근 들어 형질전환 맞추스 제작기법에 의해 $\alpha$쇄 유전자 상류에 세포특이적 발현을 조절하는 특이부위가 존재함이 보고됨을 시작, FSH$\beta$ 및 LH$\beta$쇄 유전자발현을 조절하는 특이부위 또한 가까운 시기내 발견되리라 기대된다. 한편, 성선자극 호르몬 방출호르몬(GnRH), 스테로이드 호르몬 및 여러 결합단백질과 같은 세포의 신호는 각기 다른 신호전달체계를 통하여 GTH유전자 발현을 일으킨다. 또한 뇌하수체에서도 그 존재가 확인된 전사인자들 (cFos, cJun)과 미지의 인자들은 상호간에 다양한 이량체를 형성하여 유전자 발현을 조절하는 각 특이부위에 결합함으로써 전사단계에서의 다양한 제어가 존재함이 밝혀지고 있으며 이러한 유전자상의 특이발현영역과 세포의 신호별 전사인자에 관한 연구는 번식에 있어 중요한 성선자극호르몬에 관한 분자수준의 조절기전을 밝혀내리라 기대되어진다.

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Establishment and characterization of porcine mammary gland epithelial cell line using three dimensional culture system (3차원 배양 시스템을 이용한 돼지 유선 상피 세포 주 특성과 설정)

  • Chung, Hak-Jae
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.10
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    • pp.551-558
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    • 2017
  • To study and validate tissue-specific promoters and vectors, it is important to develop cell culture systems that retain the tissue and species specificity. Such systems are attractive alternatives to transgenic animal models. This study established a line of porcine mammary gland epithelial cells (PMECs) from a primary culture based on the cellular morphology and mRNA levels of porcine beta-casein (CSN2). The selected PMECs were stained with the cytokeratin antibody, and were shown to express milk protein genes (CSN2, lactoferrin, and whey acidic protein). In addition, to confirm the acini structure of PMEC932-7 in 3D culture, live cells were stained with SYTO-13 dye, which binds to nucleic acid. The acini of these PMECs on matrigel were formed by the aggregation of peripheral cells and featured a hollow lumens. The system was demonstrated by testing the effects of the culture conditions to cell culture including cell density and matrigel methods of the PMECs. These results suggest that PMECs possess the genetic and structural features of mammary epithelial cells.

Effects of Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture on Osteoblast Differentiation and Osteoclast Formation (방사선 육종 차조기와 백출 복합물이 조골세포와 파골세포의 활성에 미치는 영향)

  • Sim, Boo-Yong;Ji, Joong-Gu
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.1
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    • pp.168-177
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    • 2021
  • The effects of the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture on the activities of osteoblast differentiation and the restraint of osteoclast formation were investigated. the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture in the human osteoblast "MG-63" cell, was examined in relation to alkaline phosphatase (ALP) activity and alizarin red stains. In order to observe the effects of osteoclasts formation, we analyzed RAW 264.7 cell tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains. In cytotoxicity testing, it was confirmed that apple extract is safe at a concentration of 50 ㎍/㎖ or less. The ALP activity and Bone calcification formation ability were the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture had a lower activity than that of control group. However the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture significantly reduced activity of TRAP in the RAW 264.7 osteoclastic cell generation and effectively Inhibited the TRAP(+) multinuclear cells. Thus, our results demonstrate that the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture enhances the inhibitory activity of bone-resorption in RAW 264.7 cells. In conclusion, the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture seem to be effective in the prevention and treatment of bone related disorders.

Expression and Production of Human Granulocyte Colony Stimulating Factor (G-CSF) in Silkworm Cell Line (누에세포를 이용한 인간 G-CSF의 발현 및 생산)

  • Park, Jeong-Hae;Jang, Ho-Jung;Kang, Seok-Woo;Goo, Tae-Won;Chung, Kyung-Tae
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1577-1581
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    • 2010
  • Granulocyte colony stimulating factor (G-CSF) is a hematopoietic cytokine that stimulates bone marrow cells to proliferate and differentiate into granulocytes. G-CSF is approved and used for therapeutic purposes. The endoplasmic reticulum (ER) signal peptide of hG-CSF was replaced with silkworm-specific signal peptides to express and efficiently secrete recombinant hG-CSF by silkworm cells. Plasmids that contain cDNAs for hG-CSF and hG-CSF fused with silkworm- specific signal peptides of prophenoloxidase activating enzyme (PPAE), protein disulfide isomerase (PDI), and bombyxin (BX) were constructed. The G-CSF protein was expressed in insect cell line BM5 and was detected by western blot analysis. The cells transfected with plasmids containing rhG-CSF genes with silkworm-specific signal sequences released mature rhG-CSF protein more efficiently than the cells transfected with pG-CSF, the plasmid containing human G-CSF gene, including its own signal sequence. The production of hG-CSF reached maximal level at four days post-transfection and remained at a high level until 7 days post-transfection. These data demonstrate that the modification of the human G-CSF mimic to insect proteins synthesized in ER greatly improves the production of the protein.

Studies on the Heterosis Breeding in Rape by Using Cytoplasmic Genetic Male Sterile lines 3. Development of Complete Cytoplasmic Genetic Male Sterile Line "MOKPO-MS" Having Improved Quality of Oil and Oil Cake (세포질.유전자적 웅성불임 계통을 이용한 유채 Heterosis 육종에 관한 연구 제3보 유채 세포질 유전자적 웅성불임 계통의 불임안전성과 유.박의 성분 개량)

  • Lee, J.I.;Kwon, B.S.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.25 no.3
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    • pp.50-58
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    • 1980
  • A cytoplasmic-genetic male sterile line(MS) in rapes of which fertility is not restored under any temperature regimes and have the better quaility of and oil cake was developed. A MS line named Mokpo-MS, which is form the cross between Tower and Isuzu and its stamens is degenerated completely by interact~g with cytoplasmic and nuclear genes was selected. This MS line was found as a complete cytoplasmic-genetic male sterile having the quality of oil and oil cake were greatly improved by introducing Zero-erucic and Zero-glucosinolate gene from Tower.

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Taxonomic significance of the leaf micromorphology in the tribe Sorbarieae (Spiraeoideae: Rosaceae) (쉬땅나무족(조팝나무아과: 장미과) 잎표피 미세형태학적 형질의 분류학적 유용성)

  • Song, Jun-Ho;Hong, Suk-Pyo
    • Korean Journal of Plant Taxonomy
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    • v.46 no.2
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    • pp.199-212
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    • 2016
  • A comparative study of leaf epidermal microstructures in the tribe Sorbarieae (Adenostoma: 3 spp., Chamaebatiaria: 1 sp., Sorbaria: 11 spp., Spiraeanthus: 1 sp.) including related genera Gillenia (2 spp.) and Lyonothamnus (2 spp.) was carried out using scanning electron microscopy (SEM) in order to evaluate their significance in taxonomy. The leaves of Adenostoma, Chamaebatiaria, and Spiraeanthus were amphistomatic, whereas Gillenia, Lyonothamnus, and Sorbaria were hypostomatic. The size range of the guard cells is $7.84-48.7{\times}5.86-38.6{\mu}m$; the smallest one was found in Sorbaria tomentosa var. tomentosa ($7.84-11.8{\times}6.84-10.5{\mu}m$), while the largest measured example was Adenostoma fasciculatum var. obtusifolium ($30.3-48.7{\times}18.8-38.6{\mu}m$). Anomocytic stomata complex were the most frequent type (rarely cyclocytic), with usually both anomocytic and actinocytic types occurring in one leaf. On the surfaces, both the adaxial and abaxial anticlinal walls of the subsidiary cells vary (e.g., straight/curved, undulate, sinuate). Four types (unicellular non-glandular trichome, stellate, glandular trichome, pustular glandular trichome) of trichomes are found in the leaves. The epicuticular wax can be divided two types: membraneous platelets (Lyonothamnus) and platelets (Sorbaria arborea var. arborea, S. arborea var. subtomentosa, S. kirilowii, S. tomentosa var. tomentosa, Spiraeanthus schrenkianus). The trichome diversity (in particular, stellate, gland) and the existence of epicuticular wax may have taxonomic significance, although the leaf epidermal micromorphological characteristics do not provide synapomorphy in this tribe. These leaf micromorphological features are most likely better understood in the Sorbarieae when used in conjunction with external morphological characters.

The systematic implications of leaf micromorphological characteristics in the tribe Neillieae (Spiraeoideae, Rosaceae) (나도국수나무족(장미과) 잎 표피 미세형태학적 형질의 계통학적 유용성)

  • Song, Jun-Ho;Hong, Suk-Pyo
    • Korean Journal of Plant Taxonomy
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    • v.47 no.3
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    • pp.222-235
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    • 2017
  • A comparative study of the leaf epidermal micromorphology in the tribe Neillieae (Neillia: 4 species, 4 varieties; Physocarpus: 5 species; Stephanandra: 2 species) was carried out using scanning electron microscopy in order to evaluate the taxonomic and systematic implications of these characteristics. The leaves of the genera Neillia and Stephanandra were hypostomatic, whereas those of P. monogynus, P. opulifolius were amphistomatic. The range of the size of the stomata is $12.02-34.39{\times}10.76-27.13{\mu}m$; the smallest was found in N. thyrsiflora (average $13.98{\times}12.43{\mu}m$; $L{\times}W$), while the largest was measured in N. gracilis (average $26.82{\times}20.67{\mu}m$; $L{\times}W$). Paracytic stomata complexes are only found in N. affinis, and the anomocytic type was most commonly found. The papillate epidermal cell type was only observed on the abaxial surfaces of P. insularis. Platelet epicuticular waxes were found on the adaxial surfaces of N. affinis and S. tanakae. Four types (unicellular non-glandular, two- to five-armed, stellate, and glandular) of trichomes were found on the leaves. Stellates were observed in all species of Physocarpus except for P. insularis. Consequently, leaf epidermal micromorphological characteristics (e.g., the presence of papillate epidermal cells and stellate, and stomata complexes) may have high taxonomic and systematic value in Neillieae. Our results strongly support previous molecular phylogenetic and palynological hypotheses that Stephanandra and Neillia are a single genus and that Physocarpus insularis should be considered as a member of Spiraea.