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이타콘산 고생산성 Aspergillus terreus 변이주의 신속 선별을 위한 효율적인 균주 스크리닝 전략 개발
신우식(Woo-Shik Shin),김평현(Pyeung-Hyeun Kim),이도훈(Dohoon Lee),김상용(Sangyong Kim),정용섭(Yong-Seob Jeong),전계택(Gie-Taek Chun) 한국생물공학회 2011 KSBB Journal Vol.26 No.3
An efficient screening method was developed for rapid selection of a few overproducers of itaconic acid (IA) among the great many mutants derived from mother strains of Aspergillus terreus. For this purpose, an attempt was made to reveal the relationships of the growth rate and sporulation of each mutant on PDA solid medium with its IA productivity in the final liquid production-culture. As a result, it was possible to classify the mutated strains into 5 groups (from [A] to [E] group) according to theirmorphologies (i.e., growth rate and sporulation extent) on the PDA slants. Notably, most of the high-yielding mutants of IA were observed to belong to [A]group which had the properties of the highest growth rate and sporulation among the 5 groups, whereas the mutant groups of [C], [D] and [E] with the contrasting morphological features showed significant reductions in their IA productivities. From these results, it was concluded that the probability of selecting IA overproducing mutants could be remarkably enhanced when the mutated colonies showing faster growth rates are firstly selected on the PDA plate, and then further screening process is performed on the basis of the sporulation extents of the mutants selected. Consequently, through the application of the strategy developed in this study, costs and time involvedin the labor-intensive task of strain improvement could be reduced to a great extent, because the time-consuming liquid culture processes did not need to performed for the unfavorable mutants belonging to the groups other than group [A].
Tissue Inhibitor of Metalloproteinases-2와 Endostatin의 혈관신생 제어 효능 평가
김수현(Soo Hyeon Kim),조영락(Young-Rak Cho),윤현재(Hyun Jae Yoon),고희영(Hee-Young Ko),김평현(Pyeung-Hyeun Kim),서동완(Dong-Wan Seo) 대한약학회 2010 약학회지 Vol.54 No.6
Therapeutic manipulation of angiogenesis, the formation of new vascular sprouts from existing capillaries, is one of the promising strategies for treatment of human diseases such as cancer, arthritis, and cardiovascular disorder. In the present study, we examined the effects and molecular mechanism of tissue inhibitor of metalloproteinases-2 (TIMP-2) and endostatin on fibroblast growth factor-2 (FGF-2)-stimulated endothelial cell proliferation, migration and adhesion in vitro, and angiogenesis in vivo. TIMP-2 and endostatin showed potent anti-angiogenic activity in vitro and in vivo. These effects appear to be mediated through different angiogenic signaling pathways. Collectively, our findings demonstrate that TIMP-2 and endostatin strongly inhibit FGF-2-induced angiogenic responses, and the establishment of fast and reproducible evaluation system in vitro and in vivo for the development of anti-angiogenic biomaterials and therapeutics.
경구면역을 통한 항원 특이적 IgA 항체 합성에 있어 Cholera Toxin과 Alginate-Microsphere의 효과
송민형,유진수,권명상,성승룡,김용희,권익찬,정서영,양재명,김평현 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.3
Secretory (S-IgA) isotype antibody (Ab) is known to play an essential role in the primary defense against various infectious agents in mucosal tissue. However, it has been mostly unsuccessful in the induction of antigen (Ag)-specific IgA Ab response in this site by peroral vaccination. In the present study, we investigated the effect of cholera toxin (CI) as a mucosal adjuvant and alginates-microspheres as a carrier on BSA-specific IgA Ab response in gut-associated lymphoid tissue`(GALT). Peroral immunization of BSA plus CT conferred a great BSA-specific IgA response but IgG response on intestinal fluid (IF). In contrast, intraperitoneal immunization of BSA with Freund's adjuvant readily induced BSA-specific IgG response but IgA response in IF. Further, number of CT specific IgA-secreting cells was substantially increased in mesenteric lymph node when CT-encapsulated-VI alginates-microspheres was administered perorally. Taken together, these results indicate that peroral immunization of soluble antigen in combination of CT or microspheres significantly enhances antigen-specific IgA response in GALT.
유전자 도입을 이용한 사람 TGF-β1 발현 세포주 제조
이종원,남상욱,김평현,이주범,전계택 한국유전학회 1999 Genes & Genomics Vol.21 No.3
Transforming Growth Factor-β1 (TGF-β1) is a pleiotrophic polypeptide involved in various biological activities including cell growth, differentiation and extracellular matrix deposition. Since TGF-β1 plays important roles in wound healing processes as well as in immune regulation, TGF-β1 is considered as a potential therapeutic agent for wound healing or organ transplantation. To obtain large quantity of human recombinant TGF-β1, an expression plasmid was constructed and transfected into Chinese hamster ovary (CHO) cells. A TGF-β1 producing clone was selected by measuring TGF-β1 levels in the medium and incubated in increasing concentrations of methotrexate up to 60 μM for the amplification of TGF-β1 cDNA in conjunction with a dihydrofolate reductase (dhfr) gene. The cell line selected in the medium containing 60 μM methotrexate produced TGF-β1 at the concentration of 12 ng/㎖ per 10^5 cells/day, which is approximately 9 fold over that of the transfected cells incubated without methotrexate. The selected cell line showed a slow growth rate and changed morphology. Overall, the established CHO cell line in this study would be useful to produce recombinant human TGF-β1.