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Production of Cloned Mice by Aggregation of Tetraploid Em
Bo-Woong Sim,Tsevelmaa Nanjidsuren,Chae-Won Park,Eun-Bi Seo,Min-Su Kim,Kye-Tae Chang,Kwan-Sik Min 한국동물생명공학회(구 한국동물번식학회) 2013 Reproductive & Developmental Biology(Supplement) Vol.37 No.2s
This study was investigated to optimize the efficiency of cloning and to produce the clone mice. Somatic cell nucleus transfer (SCNT) embryos were produced by injecting cumulus cell nucleus into enucleated B6D2F matured oocyte. Mouse chimeras can be also successfully produced with tetraploid host embryos. The fusion of mouse 2- cell embryos can be produced tetraploid embryos. Thus, we utilized the tetraploid embryos to increase the clone efficiency by aggregation with SCNT embryos of adult cumulus cells. SCNT embryos were found to be optimized for 6 hours activation with strontium (SrCl2). The cytochalasin B (CB) concentration (5 ug/ml) in the enucleation was evaluated in the efficiency of implantation sites and fetus offspring. Trichostatin A (TSA), histonedeacetylase inhibitor (HDACi), is recommended to production of clone mice, continuously exposed in 5~50 nM for 10 hours. And we also improved the SCNT implantation rate/ live offspring by co-transfer with parthenogenic embryo in uterus of the other site. Next, the aggregated SCNT were constructed by aggregation of SCNT embryo with tetraploid embryos to reduce epigenetic error in placenta. The pregnancy and implantation rates of aggregated SCNT were significantly higher than SCNT alone. The full term developmental rate in aggregated embryo was slightly higher than that of SCNT (3.57 vs 1.16). The placental weight of SCNT clone was significantly higher than that of in vitro fertilization as shown in previously reported. However, the placenta weight was almost reduced to IVF group in the aggregated SCNT. It was shown to be typical hyperplastic histology of mouse clones. But the aggregated SCNT method was useful to significantly reduce the placental weight known as general problems in cloned mice.
Sim, Bo-Woong,Min, Kwan-Sik 韓國受精卵移植學會 2014 한국동물생명공학회지 Vol.29 No.2
This study was conducted to optimize the efficiency of cloning and to produce cloned mice. The majority of cloned mammals derived by nuclear transfer (NT) die during gestation and have enlarged and dysfunctional placentas. In this study, the optimized conditions were established to produce clone mice. The parthenogenetic oocytes were activated after 6 h regardless of cytochalasin B (CB) concentration. CB treatment (2 μg/ml) was found second polar body. Lower concentration of CB was decreased the activation rate, but the second polar body was the best highly increased during 6 h incubation. The small fragments were exhibited in the 5 μg/ml treatment of CB, but it was not found in lower concentration groups (> 2.5 μg/ml). To examine effects of SrCl2 on the adult cumulus cells, somatic cell NT oocytes were exposed during 0.5, 1 and 6 hrs. The second polar body was significantly greater in 0.5 h exposure group (6.6%) than 1, 6 hrs. Developmental rate from 2-cell to 4-cell was the lowest in 7.5 mM Strontium chloride (SrCl2) groups (84.1% and 64.3%) than 5, 10 m MSrCl2. The implantation rate was not significantly difference among 5, 7.5 and 10 m MSrCl2 group. Three live fetuses were produced by SCNT. SCNT placentas were remarkably heavier than IVF group (8 fetuses) (0.34, 0.34, 0.33 vs 0.14 g) compared with the placenta weight of IVF and SCNT clones. (Key words : parthenogenetic oocytes, cytochalasin B, cloned mice)
후박군 ( Bo-jun Hou ),오외 ( Wei Wu ),장홍정 ( Hong-ting Zhang ),심영찬 ( Young-chan Sim ),김웅식 ( Woong-sik Kim ) 한국정보처리학회 2009 한국정보처리학회 학술대회논문집 Vol.16 No.2
분 논문에서는 원격 온도제어시스템을 연구하였다. 원격계측 센서를 이용하여 비닐하우스 내의 온도 정보를 수집하고 이 데이터를 무선통신을 이용하여 서버에 전송 후 온도제어 정보를 다시 히터시스템에 전달한다. 옥내에서 비닐하우스 내 온도변화를 파악하고 원격으로 히터시스템을 가동할 수 있다.
가스 조리기용 다공성 버너의 연소특성에 관한 실험적 연구
김환웅(Hwan-Woong Kim),변장희(Jang-Hee Byeon),심윤보(Yun-Bo Sim),심근선(Keun-Seon Sim),이기만(Kee-Man Lee) 한국산학기술학회 2014 한국산학기술학회 학술대회 Vol.- No.-
본 연구는 가스 조리기용 표면연소 버너중 하나인 다공성 금속 매트 버너를 열용량과 당량비에 따라 버너의 특징 및 성능을 분석하고, 다공성 버너가 가진 특징 중 하나인 미연혼합기의 예열효과에 관하 여 연구하였다. 배기가스의 온도는 전체적으로 당량비가 증가할수록 감소하는 경향을 보이며 당량비 1.0을 기준으로 다시 증가하는 특성을 나타내었다. 그런데 미연혼합기의 온도는 당량비가 증가할수록 증가하는 경향을 보이지만 각각의 최대점을 기준으로 다시 감소하는 경향을 보였다. 연소 배기성능으 로는 CO와 NOx 측정하여 비교하였고, CO의 경우 0.9 이후에는 풀(Pool) 형태의 외염이 형성되면서 급격하게 증가하는 특성을 보였다. NOx의 경우 당량비가 증가하면 같이 증가하다 당량비 1.0을 중심 으로 최대치를 보이다가 다시 감소하는 경향을 가졌다. CO와 NOx 모두 열용량이 증가할수록 최대점 은 낮은 당량비에서 나타나며 배출량은 서로 다른 양상을 나타내는 것으로 확인하였다.
TALEN-Mediated Gene Editing Method for GRK5-KO Mice
Tsevelmaa Nanjidsuren,Bo-Woong Sim,Min-Su Kim,Chae-Won Park,Eun-Bi Seo,Sun-Ok Kim,Kyu-Tae Chang,Kwan-Sik Min 한국동물생명공학회(구 한국동물번식학회) 2013 Reproductive & Developmental Biology(Supplement) Vol.37 No.2s
G protein-coupled receptor kinase 5 (GRK5) is one of the seven GRK family members whose primary function is to desensitize G protein-coupled receptors (GPCRs). In recently, GRK5 deficiency has been linked to early Alzheimer disease (AD), but the mechanism by which GRK5 deficiency may accelerate to AD pathogenesis remains elusive. The GRK5 mRNA is expressed widely in brain and peripheral tissues, with highest expression evident heart, lung, and placenta. In cellular model systems, GRK5 can phosphorylate several neuronal GPCRs including ß2-adrenergic, M2-muscarinic, secretin, angiotensin AT1, and thyroid stimulating hormone receptors. Transcription activator-like effector nucleases (TALENs) are programmable nucleases that join FokI endonucleases with the modular DNA-binding domain of TALEs and highly effective in inducing mutations at specific genome loci. TALEN-mediated mutagenesis in zygotes is a potential alternative to conventional gene targeting in mice. In the presented study, we report the generation of mice with genetic knockout of the GRK gene using TALENs. We designed TALEN vectors for exon 1, 3 and 5 of mouse GRK5 gene and tested their ability to alter the each surrogate vector in 293T cells. We prepared of mRNAs for the linearized TALEN using the mMessage mMachine T7 Ultra kit. mRNAs (4ng/μl) was injected into cytoplasm of 180 one-cell embryos. After incubation for 24 hours, the selected two-cell embryos transferred into the oviduct of seven pseudopregnant C57BL/6 mice. We confirmed the genotype of Fo mice by sequencing and T7E1 assay. We found 6 mutant mice lines (11%) from 53 newborns. We also mated 3 Fo GRK5 mutant lines with wild type mice and confirmed the genotype of the F1 progenies. All the mutations observed in Fo mice were transmitted through the germline but not all progenies (8/3, 13/4, 7/4). Taken together, TALEN-mediated mutagenesis might accelerate the creation of genetically engineered mouse models and elucidate the mechanism of AD pathogenesis using GRK5 knock out mice.