• Title/Summary/Keyword: Transgenic pigs

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Effects of Insulin, Transferrin and Selenium (ITS) on In Vitro Development of Porcine Parthenogenetic and Nuclear Transfer Embryos

  • Quan, Yan-Shi;Naruse, Kenji;Kim, Baek-Chul;Kim, Hong-Rye;Han, Rang-Xun;Choi, Su-Min;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.31 no.4
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    • pp.261-265
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    • 2007
  • Insulin, transferrin and selenium (ITS) complex is reported to improve in vitro development of oocytes and embryos. This study was carried out to investigate the effects of ITS during in vitro culture (IVC) of porcine parthenogenetic and nuclear transfer (NT) embryos on subsequent developmental capacity in vitro. The electrically activated oocytes were cultured in Porcine Zygote Medium (PZM-3) with various concentrations (0, 0.1, 0.5, and 1.0%) of ITS for 7 days. Also, the electrically activated reconstructed embryos were cultured in PZM-3 with various concentrations (0, 0.1, 0.5, and 1.0%) of ITS for 6 days. Addition of ITS to culture medium did not affect development of porcine parthenogenetic embryos in vitro. To test the effect of ITS on the in vitro development of porcine NT embryos, factorial experiments were also performed for in vitro maturation (IVM) medium (TCM-199) with or without 1% ITS and culture medium (PZM-3) with or without 0.5% ITS. Addition of 0.5% ITS to culture medium increased (p<0.05) the proportion of NT blastocysts compared with non-treated group. In contrast, addition of 1% ITS to culture medium was ineffective or had a detrimental effect. Also, addition of ITS only to maturation medium increased (p<0.05) the percentage of NT blastocysts formation compared with the control group. In conclusion, addition of ITS to IVM or IVC medium could improve subsequent blastocyst development of porcine NT embryos.

Comparison of Motility, Acrosome, Viability and ATP of Boar Sperm with or without Cold Shock Resistance in Liquid Semen at 17℃ and 4℃, and Frozen-thawed Semen

  • Yi, Y.J.;Li, Z.H.;Kim, E.S.;Song, E.S.;Kim, H.B.;Cong, P.Q.;Lee, J.M.;Park, Chang-Sik
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.2
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    • pp.190-197
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    • 2008
  • This study was designed to analyze boar sperm to compare motility, acrosome morphology, viability and ATP by various preservation methods between Duroc boar A with cold shock resistance sperm and Duroc boar B with cold shock sensitivity sperm. Semen volume, sperm concentration, motility and normal acrosome between Duroc boar A and B did not show any differences within 2 h after collection. There were no differences in sperm motility and normal acrosome between boar A and B at 1 day of preservation at $17^{\circ}C$ and $4^{\circ}C$, respectively. However, sperm motility and normal acrosome from 2 day of preservation at $17^{\circ}C$ and $4^{\circ}C$, respectively, were higher for boar A than boar B. The frozen-thawed sperm motility and normal acrosome were higher for boar A than boar B. The sperm viability and ATP concentration according to storage period of liquid semen at $17^{\circ}C$ and $4^{\circ}C$ were higher for boar A than boar B. Also, the sperm viability and ATP concentration of frozen-thawed semen were higher for boar A than boar B. In conclusion, we found out that the original quality of boar semen with cold shock resistance sperm played an important role.

Analysis of Membrane Integrity and Mitochondrial Activity in Fresh and Cryopreserved Boar Sperm Using Flow Cytometry

  • Park C. S.;Li Z. H.;Sung N. D.;Jin D. I.;Cong P. Q.;Kim E. S.;Song E. S.;Yi Y. J.
    • Reproductive and Developmental Biology
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    • v.29 no.4
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    • pp.253-257
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    • 2005
  • This study was carried out to evaluate the effects of washing medium, breed and washing temperature of fresh and frozen-thawed boar sperm on mitochondrial activity and membrane integrity by flow cytometry. More than $80\%$ of fresh sperm washed with mTLP-PVA medium at $20^{\circ}C$ exhibited an intact membrane and a functional mitochondrion. With frozen-thawed samples, a large number of sperm showed both damaged membrane $(36.4\~46.9\%)$ and nonfunctional mitochondrion $(55.1\~71.1\%)$ in the mTLP-PVA and BTS washing media at $20^{\circ}C$. There were no breed effects of fresh and frozen-thawed sperm on mitochondrial activity and membrane integrity. The percentages of damaged membrane of fresh and frozen sperm, respectively, were higher at $4^{\circ}C$ washing temperature than at $20^{\circ}C$ washing temperature in the mTLP-PVA medium. We found that washing medium and washing temperature of fresh and frozen-thawed boar sperm were important for the analyses of mitochondrial activity and membrane integrity by flow cytometry.

Increased of the Red Blood Cell in Peripheral Plasma of Transgenic Pigs Harboring hEPO Gene

  • Park, J.K.;Jeon, I.S.;Lee, Y.K.;Lee, P.Y.;Kim, S.W.;Kim, S.J.;Lee, H.G.;Han, J.H.;Park, C.G.;Min, K.S.;Lee, C.H.;Lee, H.T.;Chang, W.K.
    • Korean Journal of Animal Reproduction
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    • v.27 no.4
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    • pp.317-324
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    • 2003
  • The present study were performed to analysis the hematocrit and the red blood cells content into the blood plasma of the transgenic pigs harboring recombinent human erythropoietin gene (rhEPO). Mouse whey acidic protein (mWAP) linked to rhEPO gene was microinjected into pronuclei of porcine one-cell zygotes. After delivered of offspring, PCR analyses identified one mWAP-rhEPO transgenic founder offspring(F$_{0}$). The first generation of transgenic pig (F$_{0}$) harboring mWAP-hEPO appeared to be a male, and the second generation (F$_1$) pigs were made by natural mating of F$_{0}$ with domestic swine, and male and female transgenic pigs (F$_1$) were identified by PCR. The blood samples from transgenic and normal pigs were collected for 50 days during lactation and were counted the red blood cell (RBC) numbers and Hematocrit (HCT) content into the blood. The transgenic pigs expressing rhEPO in their blood gave rise to higher RBC numbers and HCT contents than control animals. rhEPO was secreted both in the blood and milk of genetically engineered pigs harboring rhEPO gene. Therefore, this study provides a model regarding the production of transgenic pig carrying hEPO transgene for biomedical research.earch.

Growth Regulation in IGF-1 Receptor Transgenic Mice

  • Kim Hyun-Joo;Shin Young-Min;Chang Suk-Min;Park Chang-Sik;Jin Dong-Il
    • Reproductive and Developmental Biology
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    • v.30 no.2
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    • pp.93-97
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    • 2006
  • To study the signaling effect of insulin-like growth factor-I(IGF-1), transgenic mice containing IGF-1 Receptor (IGF-1R) cDNA fused to metallothionein promoter were produced by DNA microinjection into the pronucleus of mouse zygote. Three founders were produced with transgenic mice containing IGF-1R gene. Transgenic mice lines contained approximately $4{\sim}20$ copies of transgenes per cell and transmission of this gene into the progeny with Mendelian manner were determined. The founder mice were mated with normal mice to produce $F_1$ mice and then $F_2$ mice. Transmission rates of IGF-1R transgene in the progeny mice were $25{\sim}60%$ in $F_1$ generation and $40{\sim}50%$ in $F_2$ generation. The mRNA expression of IGF-1R transgene in liver was analyzed using RT-PCR for IGF-1R gene in liver. When body weights of transgenic pups were measured during 4, 10 and 14 weeks after birth, IGF-1R transgenic mice grew faster than non transgenic littermates. This study indicated that growth regulation by IGF-1 signaling through IGF-1R can be elucidated using IGF-1R transgenic mice.

Mitochondrial DNA Diversity of Korean Ogol Chicken

  • Bhuiyan, M.S.A.;Lee, Y.J.;Chung, H.J.;Jung, W.Y.;Lee, J.H.
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2006.11a
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    • pp.77-79
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    • 2006
  • 한국재래오골계는 천연기념물로 등록이 되어 있어 세계의 중요한 유전자원 중 하나이다. 현재 한국에서 사육되어 있는 오골계의 유전적 특성을 규명하기 위하여 미토콘드리아 DNA의 변이를 이용하여 계통 분석을 실시하였다. 총 31 마리의 한국재래오골계가 이 분석에 이용되었으며 10개의 haplotypes이 관찰되었다. NJ 방법으로 만들어진 계통도 분석을 통하여 이미 닭에서 알려진 A부터 C의 lineage를 포함하는 것으로 보아 한국 재래오골계는 아직도 높은 유전적 다형성을 유지하고 있음을 알 수 있었다. 이 연구 결과는 한국 재래오골계의 육종 및 보존 계획을 세우는데 유용하게 이용될 수 있을 것으로 사료된다.

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Mitochondrial DNA Diversity of Korean Ogol Chicken

  • Lee, Y.J.;Bhuiyan, M.S.A.;Chung, H.J.;Jung, W.Y.;Choi, K.D.;Jang, B.G.;Paek, W.K.;Jeon, J.T.;Park, C.S.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.4
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    • pp.477-481
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    • 2007
  • Korean Ogol chicken has been registered as a natural monument in Korea and regarded as a valuable genetic resource for the world. As an initial step to investigate the genetic structures of this breed, phylogenetic analysis and calculation of genetic diversities have been performed using mitochondrial DNA (mtDNA) sequence variations. A total of 31 Korean Ogol chicken was grouped into four haplotypes and the large haplotype was represented in 12 individuals. The unrooted neighbor-joining tree indicates that the Korean Ogol chicken shared three (A to C) major chicken lineages representing the high genetic variability of this breed. These results can be used for making the breeding and conservation strategies for the Korean Ogol chicken.

Comparison of hematologic and biochemical values in htPA transgenic pigs (사람 조직 플라스미노겐 활성인자 생산용 형질전환 돼지에서의 혈액학적 성상 비교)

  • Park, Mi-Ryung;Hwang, In-Sul;Lee, Seunghoon;Lee, Hwi-Cheul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.12
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    • pp.395-400
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    • 2020
  • Pigs have been used widely in biomedical research owing to their physiologic and anatomic similarities to humans. Analysis of the hematologic and biochemical values in pigs is an important basis for biomedical research and veterinary clinical diagnosis, but research on transgenic pigs has been sparse. This study was conducted to obtain basic data on transgenic pigs and to describe and compare the reference values for hematologic and biochemical parameters in human tissue plasminogen activator (htPA) transgenic pigs vs normal pigs. Blood samples were obtained from 7 normal LY (Landrace-Yorkshire crossbred) pigs and 8 transgenic pigs and 16 hematologic and 15 serum biochemical parameters were tested. Among the hematologic parameters tested, significant differences were observed in the red blood cells (RBC), mean red blood cell hemoglobin (MCH), and lymphocytes (LYM), between the non-transgenic and transgenic pigs. Among the biochemical parameters tested, the blood urea nitrogen (BUN), total protein (TP), cholesterol (CHOL), alanine aminotransferase (ALT), creatinine (CREA), gamma glutamyl transpeptidase (GGT), globin (GOB), and amylase (AMYL) showed significant differences between the two groups. Thus, the values determined in this study can be used as basic reference values for transgenic pigs and will contribute to their use in biomedical research.

Activation Mechanism of Protein Kinase B by DNA-dependent Protein Kinase Involved in the DNA Repair System

  • Li, Yuwen;Piao, Longzhen;Yang, Keum-Jin;Shin, Sang-Hee;Shin, Eul-Soon;Park, Kyung-Ah;Byun, Hee-Sun;Won, Min-Ho;Choi, Byung-Lyul;Lee, Hyun-Ji;Kim, Young-Rae;Hong, Jang-Hee;Hur, Gang-Min;Kim, Jeong-Lan;Cho, Jae-Youl;Seok, Jeong-Ho;Park, Jong-Sun
    • Toxicological Research
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    • v.24 no.3
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    • pp.175-182
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    • 2008
  • DNA-dependent protein kinase(DNA-PK) is involved in joining DNA double-strand breaks induced by ionizing radiation or V(D)J recombination and is activated by DNA ends and composed of a DNA binding subunit, Ku, and a catalytic subunit, DNA-PKcs. It has been suggested that DNA-PK might be $2^{nd}$ upstream kinase for protein kinase B(PKB). In this report, we showed that Ser473 phosphorylation in the hydrophobic-motif of PKB is blocked in DNA-PK knockout mouse embryonic fibroblast cells(MEFs) following insulin stimulation, while there is no effect on Ser473 phosphorylation in DNA-PK wild type MEF cells. The observation is further confirmed in human glioblastoma cells expressing a mutant form of DNA-PK(M059J) and a wild-type of DNA-PK(M059K), indicating that DNA-PK is indeed important for PKB activation. Furthermore, the treatment of cells with doxorubicin, DNA-damage inducing agent, leads to PKB phosphorylation on Ser473 in control MEF cells while there is no response in DNA-PK knockout MEF cells. Together, these results proposed that DNA-PK has a potential role in insulin signaling as well as DNA-repair signaling pathway.

Insertional Variations of Two Porcine Endogenous Retroviruses (PERVs) in Korean Native Pigs and Asian Wild Boars

  • Jung, K.C.;Yu, S.L.;Kim, T.H.;Jeon, J.T.;Rogel-Gaillard, C.;Park, C.S.;Jin, D.I.;Moran, C.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.4
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    • pp.461-465
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    • 2007
  • Porcine Endogenous Retroviruses (PERVs) are a major concern in relation to xenotransplantation. Previous research indicated that PERVs are present at about 50 copies in the pig genome and their chromosomal insertion sites are different among pig breeds. We examined nine Korean native pigs and seven Asian Wild Boars for the presence of a PERV-A at SSC 1q2.4 and a PERV-B at SSC 7p1.1-2 previously reported in a Large White pig. The PERV-B at locus 7p1.1-2 displayed insertional variability in Korean native pigs and Asian Wild Boars. Using the primers for the PERV-A at 1q2.4 from Large White pig, we only can amplify an unclassified 798 bp sequence, which showed insertional variability only in Korean native pigs. This study indicates that there are differences within and between Asian and European pigs in PERV insertions and suggests that selection could generate PERV-free lines of pigs more suitable for xenotransplantation.