• 제목/요약/키워드: Transgenic pig

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CONSIDERATIONS IN THE DEVELOPMENT OF FUTURE PIG BREEDING PROGRAM - REVIEW -

  • Haley, C.S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제4권4호
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    • pp.305-328
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    • 1991
  • Pig breeding programs have been very successful in the improvement of animals by the simple expedient of focusing on a few traits of economic importance, particularly growth efficiency and leanness. Further reductions in leanness may become more difficult to achieve, due to reduced genetic variation, and less desirable, due to adverse correlated effects on meat and eating quality. Best linear unbiased prediction (BLUP) of breeding values makes possible the incorporation of data from many sources and increases the value of including traits such as sow performance in the breeding objective. Advances in technology, such as electronic animal identification, electronic feeders, improved ultrasonic scanners and automated data capture at slaughter houses, increase the number of sources of information that can be included in breeding value predictions. Breeding program structures will evolve to reflect these changes and a common structure is likely to be several or many breeding farms genetically linked by A.i., with data collected on a number of traits from many sources and integrated into a single breeding value prediction using BLUP. Future developments will include the production of a porcine gene map which may make it possible to identify genes controlling economically valuable traits, such as those for litter size in the Meishan, and introgress them into nucleus populations. Genes identified from the gene map or from other sources will provide insight into the genetic basis of performance and may provide the raw material from which transgenic programs will channel additional genetic variance into nucleus populations undergoing selection.

Effects of Sperm Membrane Disruption and Electrical Activation of Oocytes on In vitro Development and Transgenesis of Porcine Embryos Produced by Intracytoplasmic Sperm Injection

  • Shim, Sang Woo;Kim, Young Ha;Lee, Hoon Taek;Shim, Hosup
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권3호
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    • pp.358-363
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    • 2008
  • The intracytoplasmic sperm injection (ICSI) procedure has recently been utilized to produce transgenic animals and may serve as an alternative to the conventional pronuclear microinjection in species such as pigs whose ooplasm is opaque and pronuclei are often invisible. In this study, the effects of sperm membrane disruption and electrical activation of oocytes on in vitro development and expression of transgene green fluorescent protein (GFP) in ICSI embryos were tested to refine this recently developed procedure. Prior to ICSI, sperm heads were treated with Triton X-100+NaCl or Triton X-100+NaCl+NaOH, to disrupt membrane to be permeable to exogenous DNA, and incubated with linearized pEGFP-N1 vector. To induce activation of oocytes, a single DC pulse of 1.3 kV/cm was applied to oocytes for $30{\mu}sec$. After ICSI was performed with the aid of a micromanipulator, in vitro development of embryos and GFP expression were monitored. The chemical treatment to disrupt sperm membrane did not affect the developmental competence of embryos. 40 to 60% of oocytes were cleaved after injection of sperm heads with disrupted membrane, whereas 48.6% (34/70) were cleaved without chemical treatment. Regardless of electrical stimulation to induce activation, oocytes were cleaved after ICSI, reflecting that, despite sperm membrane disruption, the perinuclear soluble sperm factor known to mediate oocyte activation remained intact. After development to the 4-cell stage, 11.8 (2/17, Triton X-100+NaCl+NaOH) to 58.8% (10/17, Triton X-100+NaCl) of embryos expressed GFP. The expression of GFP beyond the stage of embryonic genome activation (4-cell stage in the pig) indicates that the exogenous DNA might have been integrated into the porcine genome. When sperm heads were co-incubated with exogenous DNA following the treatment of Triton X-100+NaCl, GFP expression was observed in high percentage (58.8%) of embryos, suggesting that transgenic pigs may efficiently be produced using ICSI.

돼지에서 Zoletil/Midazolam과 Zoletil/Xylazine의 2가지 병용마취에 대한 비교 연구 (Comparative Study of Two Anesthetic Combinations (Zoletil/Midazolam and Zoletil/Xylazine) in Pigs)

  • 지현철;이재연;정성목;이수진;박창식;김명철
    • 한국임상수의학회지
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    • 제27권4호
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    • pp.330-335
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    • 2010
  • 이 연구의 목적은 돼지에 있어서 zoletil/midazolam (TZM)과 zoletil/xylazine (TZX)의 2가지 병용마취를 사용하였을 때의 마취효과와 심혈관계 및 호흡에 미치는 영향을 비교하기 위하여 실시하였다. 총 8 마리의 Landrace $\times$ Yorkshire 교잡종 돼지 ($25.3{\pm}3.3\;kg$)를 사용하였으며, 각 군마다 4 마리씩 2개 군으로 실험을 실시하였다. 첫 번째 군은 xylazine과 tiletamine/zolazepam을 2 mg/kg 용량으로 근육 내 주사하였고(TZX군), 2번째 군은 midazolam 0.5 mg/kg의 용량을 정맥 내 주사 하고 tiletamine/zolazepam 2 mg/kg 용량으로 근육 내 주사하였다(TZM군). 마취시간에 대한 평가로 induction time, anesthesia time과 standing time을 각 돼지마다 측정하였으며, 마취효과에 대한 평가로 진정, 진통, 근 이완, 자세 그리고 청각반응을 점수화 하여 매 15분마다 측정하였다. 심폐기능에 대한 평가로 심박동수와 동맥혈압, 호흡수, 직장체온을 마취 전, 마취직후, 마취 후 5분, 15분, 30분, 45분 및 60분에 각각 측정하였고, 동맥혈 가스 분석을 동일시간대에 실시하였다. 실험 결과 모든 돼지에서의 마취는 성공적이었다. 2가지 병용마취 모두 부드러운 마취 유도와 적절한 운동억제 효과를 보였으나 마취효과점수에서는 TZM군이 TZX군보다 우수하였으며, TZM군이 TZX군 보다 심폐기능 및 체온에 미치는 영향이 적으며 안정적인 것을 확인하였다. 결론적으로, TZM군은 TZX군에 비하여 더욱 양호한 마취효과를 보였으며, 심폐기능에 미치는 영향은 보다 적게 나타내었다.

돼지에서 연속형 $CO_2$ 레이저와 펄스형 $CO_2$ 레이저를 이용한 피부절개시 창상치유 평가 (Comparison of Wound Healing in Porcine Skin with Continuous-Wave and Pulsed Mode $CO_2$ Laser Incisions)

  • 이재연;조성환;박창식;김명철
    • 한국임상수의학회지
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    • 제27권6호
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    • pp.647-651
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    • 2010
  • 연속형 $CO_2$ 레이저와 펄스형 $CO_2$ 레이저를 이용한 돼지 피부절개 시 창상 치유에 미치는 영향을 평가하고자 본 실험을 실시하였다. 다섯 마리의 돼지(Landrace x Yorkshire) (45-51 kg, 4-6 개월령, 수컷 3마리, 암컷 2마리)를 이용 하였고, 각각의 돼지에서 우측 및 좌측의 등쪽 피부에 대칭적으로 연속형 $CO_2$ 레이저와 펄스형 $CO_2$ 레이저를 이용하여 절개($2{\times}2{\times}2cm^2$) 하였다. 양측 피부 절개는 Maxon 3-0 를 이용하여 봉합하였다. 수술 후 3, 7, 14, 21일에 병리조직학적 검사를 실시하였다. 창상 부위의 재상피화는 연속형 $CO_2$ 레이저 군에 비해 펄스형 $CO_2$ 레이저군에서 더 많이 이루어졌다. 육아조직 형성은 창상후 경과일 3일에 펄스 $CO_2$ 레이저군에서 유의적으로 높게 나타났다(P < 0.05). 섬유아세포는 창상후 경과일 7일에 펄스형 $CO_2$ 레이져군에서 유의적으로 많게 형성되었다(P < 0.05). 결론적으로 피부절개 시에 있어서 펄스형 $CO_2$ 레이저는 연속형 $CO_2$ 레이저에 비하여 재상피화, 육아조직형성 및 섬유아세포가 더 높게 나타났으며, 레이저 시술에 따른 조직손상을 적게 나타내었다. 따라서 피부절개 시에 있어서 펄스형$CO_2$ 레이저가 연속형 $CO_2$ 레이저 보다 더 적합할 것으로 판단된다.

Characterizations of Cell Lineage Markers in the Bone Marrow Cells of Recloned GFP Pigs for Possible Use of Stem Cell Population

  • Park, Kwang-Wook;Choi, Sung-Sik;Lee, Dong-Ho;Lee, Hwang;Choi, Seung-Kyu;Park, Chang-Sik;Lee, Sang-Ho
    • Reproductive and Developmental Biology
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    • 제35권1호
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    • pp.23-31
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    • 2011
  • Two piglets and one juvenile pig were used to investigate closely what types of cells express green fluorescent protein (GFP) and if any, whether the GFP-tagged cells could be used for stem cell transplantation research as a middle-sized animal model in bone marrow cells of recloned GFP pigs. Bone marrow cells were recovered from the tibia, and further analyzed with various cell lineage markers to determine which cell lineage is concurrently expressing visible GFP in each individual animal. In the three animals, visible GFP were observed only in proportions of the plated cells immediately after collection, showing 41, 2 and 91% of bone marrow cells in clones #1, 2 and 3, respectively. The intensity of the visible GFP expression was variable even in an individual clone depending on cell sizes and types. The overall intensities of GFP expression were also different among the individual clones from very weak, weak to strong. Upon culture for 14 days in vitro (14DIV), some cell types showed intensive GFP expression throughout the cells; in particular, in cytoskeletons and the nucleus, on the other hand. Others are shown to be diffused GFP expression patterns only in the cytoplasm. Finally, characterization of stem cell lineage markers was carried out only in the clone #3 who showed intensive GFP expression. SSEA-1, SSEA-3, CD34, nestin and GFAP were expressed in proportions of the GFP expressing cells, but not all of them, suggesting that GFP expression occur in various cell lineages. These results indicate that targeted insertion of GFP gene should be pursued as in mouse approach to be useful for stem cell research. Furthermore, cell- or tissue-specific promoter should also be used if GFP pig is going to be meaningful for a model for stem cell transplantation.

Production of Cloned Korean Native Pig by Somatic Cell Nuclear Transfer

  • Hwang, In-Sul;Kwon, Dae-Jin;Oh, Keun Bong;Ock, Sun-A;Chung, Hak-Jae;Cho, In-Cheol;Lee, Jeong-Woong;Im, Gi-Sun;Hwang, Seongsoo
    • 한국발생생물학회지:발생과생식
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    • 제19권2호
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    • pp.79-84
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    • 2015
  • The Korean native pig (KNP) have been considered as animal models for animal biotechnology research because of their relatively small body size and their presumably highly inbred status due to the closed breeding program. However, little is reported about the use of KNP for animal biotechnology researches. This study was performed to establish the somatic cell nuclear transfer (SCNT) protocol for the production of swine leukocyte antigens (SLA) homotype-defined SCNT KNP. The ear fibroblast cells originated from KNP were cultured and used as donor cell. After thawing, the donor cells were cultured for 1 hour with 15 ${\mu}M$ roscovitine prior to the nuclear transfer. The numbers of reconstructed and parthenogenetic embryos transferred were $98{\pm}35.2$ and $145{\pm}11.2$, respectively. The pregnancy and delivery rate were 3/5 (60%) and 2/5 (40%). One healthy SLA homotype-defined SCNT KNP was successfully generated. The recipient-based individual cloning efficiency ranged from 0.65 to 1.08%. Taken together, it can be postulated that the methodological establishment of the production of SLA homotype-defined cloned KNP can be applied to the generation of transgenic cloned KNP as model animals for human disease and xenotransplantation researches.

Seroprevalence of selected porcine respiratory pathogens in the pig herds in Chungcheong and Gyeongsang provinces in Korea

  • Chu, Jia-Qi;Hu, Xu-Min;Kim, Myung-Cheol;Park, Chang-Sik;Jun, Moo-Hyung
    • 한국동물위생학회지
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    • 제32권1호
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    • pp.27-32
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    • 2009
  • We studied the seroprevalence of four respiratory pathogens in Korean swine farms located in Chungnam, Chungbuk, Gyeongnam and Gyeongbuk provinces during the period of spring of 2007 to winter of 2008. Serological tests were performed using commercial ELISA kits. A total of 530 serum samples were tested for the antibodies against porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), Mycoplasma hyopneumoniae (M. hyo) and Actinobacillus pleuropneumoniae (APP). Seroprevalence for four respiratory pathogens were estimated by ELISA-positive rates of the submitted samples. The overall seropositive rates of PRRSV, APP, M. hyo and PCV2 were 32.6%, 10.6%, 38.4% and 88.5%, respectively. By production stage, the seropositive rate for PRRSV was highest in nursery pig populations (46.2%). In contrast, the highest seropositive rates of APP and M. hyo were observed in sow and growing pigs. However, the seroprevalence of PCV2 was ranged from 85.7% to 89.6%, showing no significant difference among the production stages. In the seroprevalence by season, PRRSV, APP and M. hyo infections revealed typical seasonal patterns that the peaks of the seropositive rates were observed between early winter and late spring. In case of PCV2, no particular seasonal patterns were noticed. The pig herds in Gyeongbuk province where PMWS was endemic during the period of survey showed the highest seropositive rates for PRRSV (44.6%), M. hyo (47.5%), and PCV2 (92.7%). Seropositive rates for APP of four provinces were approximately 10%. These results might be valuable for control and prevention of the respiratory diseases and helpful to define strategies related to vaccine applications.

Embryonic Stem Cell and Nuclear Transfer

  • 임정묵
    • 한국수정란이식학회:학술대회논문집
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    • 한국수정란이식학회 2002년도 춘계학술세미나 및 워크숍
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    • pp.19-25
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    • 2002
  • Researches on manipulation pluripotent stem cells derived from blastocysts or promordial germ cells (PGCs) have a great advantages for developing innovative technologies in various fields of life science including medicine, pharmaceutics, and biotechnology. Since the first isolation in the mouse embryos, stem cells or stem cell-like colonies have been continuously established in the mouse of different strains, cattle, pig, rabbit, and human. In the animal species, stem cell biology is important for developing transgenic technology including disease model animal and bioreactor production. ES cell can be isolated from the inner cell mass of blastocysts by either mechanical operation or immunosurgery. So, mass production of blastocyst is a prerequisite factor for successful undertaking ES cell manipulation. In the case of animal ES cell research, various protocol of gamete biotechnology can be applied for improving the efficiency of stem cell research. Somatic cell nuclear transfer technique can be applied to researches on animal ES cells, since it is powerful tool for producing clone embryos containing genes of interest. In this presentation, a brief review was made for explaining how somatic cell nuclear transfer technology could contribute to improving stem cell manipulation technology.

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Recent Development in Embryo Technology in Pigs - Review -

  • Niwa, K.;Funahashi, H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제12권6호
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    • pp.966-975
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    • 1999
  • Technologies on preimplantation porcine embryos have been developed quickly and significantly. Successful development of systems for culture of porcine zygotes to the blastocyst stage has made it possible to utilize follicular oocytes for in vitro production of embryos and thus stimulated research on various embryo technologies. Recent technological development of embryo cryopreservation, separation of X- and Y-bearing spermatozoa and non-surgical embryo transfer has also made it easy to utilize in vivo- and in vitro-produced embryos for artificial manipulation to produce clones and transgenic pigs. Further progress in overcoming various problems associated with each embryo technology will result in acceptable efficiency to utilize porcine embryos with a high or increased quality. Combining these technologies will accelerate further expansion of the swine industry not only for meat production but also for the production of therapeutic recombinant proteins and xonografts.

Catalytic Hydrolysis of Phosphate Diesters as DNA Model with Tetranuclear Nickle (II) Complex

  • Sung, Nack-Do;Kim, Tae-Young
    • Journal of Applied Biological Chemistry
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    • 제49권3호
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    • pp.86-89
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    • 2006
  • The novel tetranuclear nickel (II) complex is a high rate accelerator in promoting hydrolysis of phosphate diesters. Nickel-bound bis-nitrophenyl phosphate (BNPP) can be $10^4$ times more reactive than the unbound BNPP. The large rate of enhancements by the complex slightly under basic condition has shown high catalytic activity in phosphate diester cleavage. The bell-shaped pH-rate profile indicated that the nickel-oxide form of the tetranuclear complex or its kinetic equivalent was the active species for cleaving BNPP. The catalytic hydrolysis between tetranuclear nickel (II) complex and phosphate diester proceeds via the formation of bidentate coordination of the anionic phosphate to the Ni (II) atom. This reveals that the complex has the possibility as artificial nuclease.