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Identification of a putative transactivation domain in human Nanog
오종현,도현진,양흥모,문신영,차광율,정형민,김재환 생화학분자생물학회 2005 Experimental and molecular medicine Vol.37 No.3
Nanog is a newly identified divergent homeodo-main protein that directs the infinite propagation and sustains the pluripotency of embryonic stem cels. It has been reported that murine Nanog has two potent transactivation domains in N-terminal and C-terminal regions. Human Nanog (hNanog) to the open reading frame and homeodomain of murine Nanog, respectively. However, the func-tional domains and molecular mechanisms of hNanog are poorly understood. In this study, for the first time, we presented that only C-terminus of hNanog contains a potent transactivation do-main. Based on the amino acid sequences of homeobox domain, we roughly divided hNanog open reading frame into the thre regions such as N-terminal, homeodomain and C-terminal re-gions and constructed either the fusion pro-binding domain or the context of native hNanog protein. Reporter asays by using reporter pla-mid containing Gal4 or Nanog binding site revealed that the only C-terminal region exhibited the significant fold induction of transactivation. However, interestingly, there was no significant activation through N-terminal region unlike mu-rine Nanog, suggesting that C-terminal region may have more critical roles in the transcriptional activation of target genes. Taken together, the finding of a putative transactivation domain in hNanog may contribute to the further understand-downstream genes involved in self-renewal and pluripotency of human stem cels.
초자화 냉동법으로 냉동.해동한 Neonatal 생쥐 난소의 생체내 동소이식 후 난포 발달에 관한 연구
이경아,이숙현,윤세진,고정재,차광열,Lee, K.A.,Lee, S.H.,Yoon, S.J.,Ko, J.J.,Cha, K.Y. 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.2
Ovarian development of the vitrified neonatal ovaries after orthotopical transplantation into the ovariectomized adult recipient mouse were observed. Ovaries were collected from the neonatal females on day of birth and grouped for fresh, vitrification for 1-minute, and 3-minute. Vitrified and thawed neonatal ovaries were orthotopically transplanted into ovarian bursa of the adult mice from which endogenous ovaries have removed just prior to the transplantation (1 minute: n=25; 3 minutes n=23). Fresh ovarian tissue transplanted (n=25) mice were included as control groups. Returning of the estrus cycles and the survival and development of the transplanted ovaries were evaluated. Intact ovaries from neonatal, and four weeks old mice were used for comparison of the ovarian development as in vivo-developed control. From 2 weeks after transplantation, 64%, 36%, and 75% of the transplanted mice showed return of the estrus cycles in fresh, 1-minute, and 3-minute groups, respectively. Four weeks after transplantation, all mice were sacrificed and ovarian tissues were recovered for histological analysis. 57.1%, 33.3%, and 64.7% mice in fresh, 1-minute, and 3-minute groups, respectively, had survived ovaries with follicles at various stages of growth from primordial to preovulatory follicles. Corpus lutea were also observed. Results of the present study suggest that 1) normal folliculogenesis has initiated in vivo after vitrification, and 2) the vitrification may be used as a preservation method for ovarian tissues for establishment of ovarian tissue bank.
구송이,차광현,송대근,이동언,판철호 한국응용생명화학회 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.5
High hydrostatic pressure (HHP) treatment was used to increase the concentration of sulforaphane in Brussels sprout, a cruciferous vegetable known to have health benefits. The highest concentration of sulforaphane was 1021.8 μmol per kg fresh weight of Brussels sprouts after HHP treatment at 500 MPa,which corresponded to a 317% increase compared to the HHPuntreated control.
이경아,이숙현,하상덕,윤세진,고정재,이우식,윤태기,차광열,Lee, K.A.,Lee, S.H.,Ha, S.D.,Yoon, S.J.,Ko, J.J.,Lee, W.S.,Yoon, T.K.,Cha, K.Y. 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.2
The present study was conducted to evaluate whether vitrification could be used for ovarian tissue preservation. The important issue here is that the vitrification is very simple, easy, and economical compared to the conventional cryopreserving method that using automatic freezing instrument. Human ovarian cortical tissues were cryopreserved by vitrification with 5.5 M ethylene glycol and 1.0 M sucrose as cryoprotectant. Three points of temperature ($4^{\circ}C$, room temperature, and $37^{\circ}C$) and two points of duration (5 or 10 minutes) for cryoprotectant treatment were examined to determine the best condition for vitrification of the human ovarian cortical tissues. After thawing, viability of the isolated primordial follicles was examined by dye-exclusion method. Histological appearance of tissues before and after the cryopreservation was evaluated. There was no toxic effect of the 5.5 M ethylene glycol on the primordial follicles. However, when the tissues were treated with cryoprotectant at $37^{\circ}C$ for 10 minutes and exposed to liquid nitrogen, it seems likely that there is certain deleterious effects on the viability of the primordial follicles. The highest viability of the primordial follicles was obtained with the treatment of cryoprotectant at room temperature for 10 minutes. Follicles and oocytes survived after freezing and thawing had the similar normal shapes as was seen in the specimens before cryopreservation. It would be useful to apply vitrification in establishing ovarian tissue banking for clinical purposes.
이은하,차광현,Trang Thi Vuong,김상민,판철호 한국응용생명화학회 2018 Applied Biological Chemistry (Appl Biol Chem) Vol.61 No.4
This study aimed to determine the bioaccessibility of lutein in lutein-rich food, using static and dynamic models of in vitro gastrointestinal digestion. Here, kale powder (KP) and lutein supplement (LS) were used as representative lutein-rich foods. The bioaccessibility of lutein from KP did not considerably differ between static (59.92%) and dynamic (56.08%) digestion. Bioaccessibility was consistently maintained at the same level during dynamic digestion. The amount of lutein released from the LS during dynamic digestion was five times higher than that released during static digestion (67.88 vs 12.34%). The results showed that (a) bioaccessibility of lutein was affected by various factors such as food source, solid:liquid ratio, and interaction with dietary components, and (b) dynamic digestion should be suitable for evaluating the bioaccessibility of lutein in high-fat foods.
서진호,임형권,차광현,정경환,Myoung-Dong Kim,이태희 한국생물공학회 2004 Biotechnology and Bioprocess Engineering Vol.9 No.6
Recombinant Saccharomyces cerevisiae expression systems were developed to produce a novel human anti-angiogenic protein called LK8, an 86 amino-acid kringle fragment protein with three disulfide linkages. Galactose-inducible LK8 expression plasmid was constructed, and LK8 production levels by four S. cerevisiae strains were compared in order to select an optimal host strain. S. cerevisiae 2805 was the most efficient among the strains tested. Elevating the LK8 gene copy number through multiple integration using -sequences as target sites resulted in more than a two-fold increase in the LK8 production level compared with the plasmid-based expression system. The maximum LK8 protein concentration of 25 mg/L was obtained from batch cultivation of the yeast transformant that harbors 16 copies of the LK8 gene. In conclusion, the strain integrated with the multiple LK8 gene secreted the protein with relatively high yield, although, the increased LK8 gene dosage over 11 copies did not lead to further enhancement in batch cultivations.
Overexpression of SOX9 in mouse embryonic stem cells directs the immediate chondrogenic commitment
김재환,도현진,양흥모,오종현,최성준,김동규,차광율,정형민 생화학분자생물학회 2005 Experimental and molecular medicine Vol.37 No.4
Mouse embryonic stem (mES) cells are capable of undergoing chondrogenesis in vitro. To en-hance this proces, the human SOX9 (hSOX9) cDNA was delivered into mES cels and the clones overexpressing hSOX9 (denoted as mES-hSOX9 cells) were verified by Western blot analysis. The grecan and Pax1 were expressed in mES-hSOX9 cells grown on feeder layers, sugesting the im-mediate efect of exogenous SOX9 on chondro-genesis. However, SOX9 overexpression did not afect the cel cycle distribution in undiferentia-ted mES cells. Upon diferentiation, colagen IIB (an adult form) was detected in day 3 imature embryoid bodies. In adition, the overexpression of exogenous SOX9 significantly induced trans-criptional activity driven by SOX9 binding site. Taken together, we for the first time demon-enous SOX9 in undiferentiated mES cels might have dual potentials to induce both chondrogenic comitment and growth capacity in the undif-ferentiated status.
임태환,이기욱,김련희,차광현,구송이,문은채,황금택 한국식품과학회 2022 Food Science and Biotechnology Vol.31 No.8
Blood trimethylamine-N-oxide (TMAO) has been associated with cardiovascular disease. Black raspberry (Rubus occidentalis, BR) has been regarded to be beneficial for cardiovascular health. This study aimed to investigate how BR extract affects serum lipid profile, gut microbial composition, metabolites in rats fed TMAO with a high-fat diet. Dietary TMAO increased serum LDL cholesterol, while BR extract decreased its level. α-Diversity of gut microbiota was not changed; however, in the rats fed TMAO, Macellibacteroides and Mucispirillum were enriched, while Ruminococcaceae was reduced. The BR supplementation could restore Macellibacteroides, Clostridium, and Ruminococcaceae. The BR supplementation increased cecal hippuric acid and serum farnesoid X receptor-antagonistic bile acids, including ursodeoxycholic acid (UDCA), tauro-α-muricholic acid, and tauro-UDCA. The BR supplementation tended to upregulate Cyp7a1 and Abcg5 expressions while downregulating Srebf2 and Hmgcr expressions. BR extract affects the gut bacterial community and microbial metabolites, lowering serum LDL cholesterol in rats with elevated serum TMAO.