• Title/Summary/Keyword: PCR condition

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Adaptive Transition of Aquaporin 5 Expression and Localization during Preimplantation Embryo Development by In Vitro Culture

  • Park, Jae-Won;Shin, Yun Kyung;Choen, Yong-Pil
    • Development and Reproduction
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    • v.18 no.3
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    • pp.153-160
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    • 2014
  • Adaptive development of early stage embryo is well established and recently it is explored that the mammalian embryos also have adaptive ability to the stressful environment. However, the mechanisms are largely unknown. In this study, to evaluate the possible role of aquaporin in early embryo developmental adaptation, the expression of aquaporin (AQP) 5 gene which is detected during early development were examined by the environmental condition. To compare expression patterns between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP5 by whole mount immunofluorescence. At in vivo condition, Aqp5 expressed in oocyte and in all the stages of preimplantation embryo. It showed peak at 2-cell stage and decreased continuously until morula stage. At in vitro condition, Aqp5 expression pattern was similar with in vivo embryos. It expressed both at embryonic genome activation phase and second mid-preimplantation gene activation phase, but the fold changes were modified between in vivo embryos and in vitro embryos. During in vivo development, AQP5 was mainly localized in apical membrane of blastomeres of 4-cell and 8-cell stage embryos, and then it was localized in cytoplasm. However, the main localization area of AQP5 was dramatically shifted after 8-cell stage from cytoplasm to nucleus by in vitro development. Those results explore the modification of Aqp5 expression levels and location of its final products by in vitro culture. It suggests that expression of Aqp5 and the roles of AQP5 in homeostasis can be modulated by in vitro culture, and that early stage embryos can develop successfully by themselves adapting to their condition through modulation of the specific gene expression and localization.

Enzyme Activity and Gene Expression of Cytochrome P450 Involved in Capsidiol Biosynthesis in Solanaceae Plants (가지과식물에서 Capsidiol 생합성에 관여하는 Cytochrome P450 유전자의 발현과 효소활성)

  • Kwon, Soon-Tae;Hasegawa, Paul
    • Korean Journal of Plant Resources
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    • v.21 no.2
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    • pp.139-143
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    • 2008
  • Enzyme activity and expression of cytochrome P450 gene involved in the pathway of capsidiol biosynthesis were compared in five different solanaceae plants such as red pepper, green pepper, tobacco, potato and egg plant. Base on genomic DNA and/or RT-PCR results, four solanaceae plants such as red pepper, green pepper, tobacco and egg plant possess P450 gene in the genome and specifically expressed by elicitor treatment. However, potato was appeared to have neither P450 nor cyclase gene in the genome. P450 genes did not show any expression in the plants under normal condition, but showed highly specific expression under elicitation condition in various organs and tissue such as leaf, root, stem and culture cells.

Gene Expression of the In Vitro Fertilized or Somatic Cell Nuclear Transfer Embryos Cultured in Medium Supplemented with Different Proteins or Energy Substrates

  • Jang, Goo;Ko, Kyeong-Hee;Jeon, Hyun-Yong;Lee, Byeong-Chun
    • Journal of Embryo Transfer
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    • v.25 no.2
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    • pp.117-125
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    • 2010
  • Several cloned animals have been produced using somatic cell nuclear transfer (SCNT) and have interested in producing the transgenic cloned animals to date. But still its efficiency was low due to a number of reasons, such as sub-optimal culture condition, aberrant gene expression and nuclear reprogramming. The purpose of this study was to analyze gene expression pattern in in vitro fertilized (IVF) or SCNT pre-implantation embryos. IVF- or SCNT-embryos were cultured in media supplemented with different proteins (FBS and BSA) or energy sources (glucose or fructose). Blastocysts from IVF or SCNT were analyzed using semi-quantitative RT-PCR in terms of developmentor metabolic-related genes. Culture medium supplemented different proteins or energy sources had affected on the expression of developmental or metabolic genes in the SCNT blastocysts.

Enhancement of Osteogenic Differentiation by Combination Treatment with 5-azacytidine and Thyroid-Stimulating Hormone in Human Osteoblast Cells

  • Sun, Hyun Jin;Song, Young Shin;Cho, Sun Wook;Park, Young Joo
    • International journal of thyroidology
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    • v.10 no.2
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    • pp.71-76
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    • 2017
  • Background and Objectives: The role of thyroid-stimulating hormone (TSH) signaling on osteoblastic differentiation is still undetermined. The aim of this study was to investigate the effects of 5-aza-2'-deoxycytidine (5-azacytidine) on TSH-mediated regulations of osteoblasts. Materials and Methods: MG63, a human osteoblastic cell-line, was treated with 5-azacytidine before inducing osteogenic differentiation using osteogenic medium (OM) containing L-ascorbic acid and ${\beta}$-glyceophosphate. Bovine TSH or monoclonal TSH receptor stimulating antibody (TSAb) was treated. Quantitative real-time PCR analyses or measurement of alkaline phosphatase activities were performed for evaluating osteoblastic differentiation. Results: Studies for osteogenic-related genes or alkaline phosphatase activity demonstrated that treatment of TSH or TSAb alone had no effects on osteoblastic differentiation in MG63 cells. However, treatment of 5-azacytidine, per se, significantly increased osteoblastic differentiation and combination treatment of 5-azacytidine and TSH or TSAb in the condition of OM showed further significant increase of osteoblastic differentiation. Conclusion: Stimulating TSH signaling has little effects on osteoblastic differentiation in vitro. However, in the condition of epigenetic modification using inhibitor of DNA methylation, TSH signaling positively affects osteoblastic differentiation in human osteoblasts.

Monocrotophos poisoning in wild mallards (Anas platyrhynchos)

  • Shim, Hang-Sub;Kim, Hae-Sung;Woo, Jong-Tae;Kim, In-Seop;Jung, Hae-Sun;Song, Eun-Ah;Bark, Jun-Jo
    • Korean Journal of Veterinary Service
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    • v.30 no.4
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    • pp.545-550
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    • 2007
  • The toxicity of organophosphate arises from disruption of the nervous system due to the inhibition of cholinesterase enzymes, leading to death. Six dead mallards were found at Ansung where is one of the most popular wintering sites for migratory birds in Korea, and requested for diagnosis to Gyeonggi Veterinary Service on January of 2007. Some examinations including polymerase chain reaction (PCR) could not find any evidence of specific disease condition. However, the contents of gastrointestinal tracts of the birds contained residues of monocrotophos ranged from 31.3ppm to 294.3ppm by gas chromatography and mass spectrometry. It can be supposed that monocroptophos was responsible for the death of mallards by this results.

DNA Marker Traceability Techniques for Hanwoo Industry

  • Lee, Jea-Young;Bae, Jung-Hwan;Lee, Yoon-Seok;Yeo, Jung-Sou
    • Journal of the Korean Data and Information Science Society
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    • v.17 no.4
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    • pp.1041-1051
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    • 2006
  • The effectiveness of genetic markers on Hanwoo traceability systems was applied and evaluated from Korean 33 Hanwoo elite sire families. Five microsatellite markers were selected finally, which were located on chromosomes different chromosomes with the end sequencing of 100 HW-YUBAC that were recorded in the NCBI by Yeungnam University. Eleven major microsatellite markers were selected from allele amplified, their frequencies, H(Heterozygosity) and PIC(Polymorphism information content) with Hardy-Weinberg equilibrium. Next, in order to evaluate the power of the markers selected on the individual animal identification with experimental condition, the match probability(MP) and the relatedness coefficient(R) were computed.

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First Case of Canine Infection with Hepatozoon canis (Apicomplexa: Haemogregarinidae) in the Republic of Korea

  • Kwon, Seung-Joo;Kim, Yoon-Hee;Oh, Hyun-Hee;Choi, Ul-Soo
    • Parasites, Hosts and Diseases
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    • v.55 no.5
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    • pp.561-564
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    • 2017
  • This report describes a dog infected with Hepatozoon canis, the first canine infection in the Republic of Korea. A 2-year-old intact male Maltese dog presented with anorexia and depression. Physical examinations revealed mild dehydration and hyperthermia ($39.8^{\circ}C$), and blood analysis showed pancytopenia. Diff-Quik staining of blood smear specimens showed the presence of ellipsoidal shaped structures (gamonts of H. canis) within a small number of neutrophils. Real-time PCR analysis using whole blood confirmed infection by H. canis. The clinical condition of the dog improved after symptomatic treatment and administration of doxycycline. Although a molecular epidemiologic survey in Korea showed H. canis infection of dogs, to our knowledge this is the first report of a dog infection in Korea molecularly shown to be H. canis.

Absence of AVP1 transcripts in wild type watermelon scions grafted onto transgenic bottle gourd rootstocks

  • Kim, Byung Oh;Han, Jeung-Sul;Park, Kyung Il;Jeon, Su Min;Kim, Chang Kil
    • Journal of Plant Biotechnology
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    • v.42 no.1
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    • pp.13-18
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    • 2015
  • In this study we confirmed the stable integration of Arabidopsis AVP1 in the genomes of bottle gourd $T_3$ homozygous lines and its transcription, and additionally evaluated possibility of translocation of the AVP1 mRNA from transgenic bottle gourd rootstocks to wild type watermelon scions. Each AVP1 gene in two bottle gourd T3 lines is abundantly expressed under a field condition. Given the grafting between wild type watermelon scions and AVP1-expressing bottle gourd rootstocks, no translocation of the AVP1 mRNA was detected in leaves, both sexual flowers, and fruits of the scions.

Humeral Hemimelia in a Holstein Calf (유우 송아지의 선천성 상완골 형성 부전)

  • 한동운
    • Journal of Veterinary Clinics
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    • v.20 no.3
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    • pp.419-422
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    • 2003
  • Humeral hemimelia is a rare congenital anomaly characterized by deficiency of the humerus with relatively intact scapular. The holstein calf was healthy expecting the deformity of forelimb humerus. The examination of PCR and serum neutralizing test was carried about BVD, akabane virus, Ibaraki virus and Kasba virus in cattle. The results were all negative. Affected calf showed forelimb deformation. Other abnormalitis was not observed in brain, spinal cord, liver, kidney and other visceral organs. Amelia are very rare limb anomalies. We report on a Holstein calf with forelimb deficiencies. This case appears humeral anomalies with ulnar and radius aplasia in calf. The condition in these calves was considered the result of a recurrence of a genetic mutation affecting a putative hemimelia locus.

Expression of the Glyphosate Resistant Gene, cp4-epsps, through Plastid Transformation in Rice (Oryza sativa L.) (벼 색소체 형질전환을 이용한 글리포세이트 저항성 유전자 cp4-epsps의 발현)

  • Kang, Kyung-Su;Kim, Min-Kyun
    • Journal of Plant Biotechnology
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    • v.33 no.2
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    • pp.75-84
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    • 2006
  • Heteroplasmic rice plastid transformant was generated using suspension cells as bombardment materials. PCR analyses confirmed incorporation of aadA and cp4-epsps genes into the rice plastid genome by homologous recombination events via the flanking sequences of the trnI and trnA. Transplastomic calli were actively proliferated when cultured on AAM2 medium supplemented with various concentrations (500-3000 mg/L) of streptomycin in dark condition, and transplastomic suspension cells showed resistance to nonselective herbicide, glyphosate. Through 'agarose pie selection' method, heteroplastomic calli, containing considerably high level of transplastome and expressing the CP4 EPSPS protein, were obtained. They were further regenerated to green shoots with healthy roots.