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      • KCI등재

        Prostaglandin A₂-induced Apoptosis is Not Inhibited by Heme Oygenase-1 in U2OS Cells

        Kyoung-Won Ko(고경원),Sun-Young Lee(이선영),Ji-Hyun Ahn(안지현),Jaetaek Kim(김재택),In-Kyung Kim(김인경),Ho-Shik Kim(김호식) 한국생명과학회 2008 생명과학회지 Vol.18 No.11

        Prostaglandin A₂ (PGA₂)는 사람 골육종 세포인 U2OS 세포주에서 apoptosis와 heme oxygenase (HO)-1의 발현을 함께 유도하였다. PGA₂에 의한 apoptosis는 HO-1의 과도한 발현이나 HO-1에 대한 small interfering RNA에 의한 발현저하에 의하여 변동되지 않았으나 H₂O₂에 의한 세포사망은 HO-1의 발현 수준에 반비례하여 변동되었다. 또한 thiol antioxidant인 N-acetyl-L-cysteine (NAC)은 PGA₂에 의한 세포사망과 HO-1의 발현 증가를 모두 차단하였지만, non-thiol antioxidant인 butylated hydroxyanisole (BHA)과 ascorbic acid는 세포사망과 HO-1의 발현 유도를 차단하지 않았다. 이와 같은 결과들은 PGA₂는 산화성 손상에 의해서가 아니라 PGA₂의 thiol-reactivity에 의하여 apoptosis와 HO-1의 발현을 유도하며, HO-1의 발현은 PGA₂에 의한 apoptosis와는 독립적인 현상이거나 기능적으로 apoptosis 유도의 하부에 위치하고 apoptosis의 진행에는 기여하지 않을 것이라는 것을 시사해 준다. Prostaglandin A₂ (PGA₂), one of cyclopentenone PGs, induced both apoptosis and heme oxygenase (HO)-1 expression in U2OS cells. PGA₂-induced apoptosis was not perturbed by either over-expression or knock-down of HO-1, whereas H₂O₂-induced cell death was inversely modulated by the expression level of HO-1. In addition, N-acetyl-L-cysteine (NAC), a thiol antioxidant, blocked both apoptosis and HO-1 expression induced by PGA₂. But, non-thiol antioxidants like butylated hydorxyanisole (BHA) and ascorbic acid did not block either apoptosis or HO-1-induction. Taken together, these results suggest that PGA₂ induces both apoptosis and HO-1 expression, which are critically related to the thiol-reactivity of PGA₂, but not oxidative stress, and HO-1 expression may be independent or functionally located downstream of apoptosis by PGA₂ without contribution to apoptosis progression.

      • SCOPUSKCI등재

        고등식물 형질전환용 유전자 운반체 pKCHI 의 개발

        정상호(Sang Ho Chung),고경원(Kyung Won Ko),홍주봉(Choo Bong Hong),최인성(In Sung Choi),정태화(Tae Wah Chung) 한국식물학회 1989 Journal of Plant Biology Vol.32 No.1

        We have developed a plasmid vector, pKCHl, for the purpose of higher plant transformation. It contains the promoter region of cauliflower mosaic virus 35S transcript (P_35s) and the terminator region of nopaline synthase gene (Tnos) with unique cloning site, Bam HI and Xba Ⅰ, between them. After inserting a foreign gene at the cloning site, P_35s-foreign gene-Tnos cassette can be recovered by using a restriction enzyme Hind Ⅲ.

      • Stephen King의 공포소설에 나타난 여성성

        송인갑 ( In-gap Song ),고경원 ( Kyung-won Ko ) 인하대학교 교육연구소 2003 교육문화연구 Vol.9 No.-

        Stephen King is well known as a popular horror writer. His works have been neglected and even ignored as mass entertainment by academic circles. However, he is reappraised as a feminist horror writer because his recent works deal with women's problems. The aim of this paper is to study the femininity in his horror novels. Also this paper investigates what King's perspective of women is and how his feminism is embodied in his four novels, Carrie (1974), Rose Madder (1995), Gerald's Game (1992) and Dolores Claiborne (1992). King creates terrible female characters in his horror novels: Carrie of Carrie, Rosie of Rose Madder, Jessie of Gerald's Game and Dolores of Dolores Claiborne. In Carrie, his protagonist is repressed by her religious fanatic mother and is alienated from her friends. Therefore, she hurts and kills them by exercising her psychokinesis. In Rose Madder, King shows the Dionysian power of a woman warrior who fights against men's violence. In Gerald's Game and Dolores Claiborne, women fight against male sexual harassment in the patriarchal society. In King's novels, the aliens who most frighten and horrify us are the women who live with and within us, yet remain isolated from a male-centered culture. Women that King's novels seem to show us are often "the perpetrators and victims of an American domestic horror". These women hide theirs alien and terrifying face beneath a familiar mask. Thus King's women in his horror novels make males fear women who live in the patriarchal society. The theory of King's domestic horror is based on Freud's "uncanny" strangeness and "otherness" of the Kristeva's horror world. The uncanny strangeness and otherness of the horror world represent the human aspects of a fanatic Dionysian world which is the opposite of the rational Apollonian. In the Dionysian world, everything is contradictory and all humans are Janus-faced. King's women prove that all human beings, not only women but also men, can reveal fanatic Dionysian power. King's terrible female characters embody the idea that it is necessary to maintain an equal and harmonious relationship between men and women in the society which does not repress women's self.

      • KCI등재

        아미노-말단 리보플라빈 생성효소 단백질의 형광 특성

        김류련,이정환,남기석,고경원,이찬용,Kim, Ryu-Ryun,Yi, Jeong-Hwan,Nam, Ki-Seok,Ko, Kyung-Won,Lee, Chan-Yong 한국미생물학회 2011 미생물학회지 Vol.47 No.1

        리보플라빈 생성효소(riboflavin synthase)는 기질인 두 분자의 6,7-dimetyl-8-ribityllumazine과 결합 후, 4-탄소 단위(4-carbon unit)의 자리 옮김 반응을 거쳐 한 분자의 리보플라빈과 한 분자의 pyrimidine 유도체를 형성하는 반응을 촉매한다. 대장균(Escherichia coli) 리보플라빈 생성효소의 아미노-말단 도메인 절반(N-terminal domain half)과 카복시-말단 도메인 절반(C-terminal domain half)은 매우 유사한 내부 자체 아미노산 서열(intra-molecular amino acid sequence)을 갖는다. 아미노-말단 영역 리보플라빈 생성효소(N-RS) 단백질의 구조와 형광 특성을 알아보기 위하여 중합효소 연쇄 반응과 위치지정 돌연변이를 통하여 10개 이상의 돌연변이 아미노-말단 리보플라빈 생성효소 단백질을 코드 하는 유전자를 증폭시켜 pQE30 벡터에 삽입한 재조합 플라스미드를 제조하여, 과발현시킨 후 분리 정제하였다. 대부분의 아미노-말단 도메인 리보플라빈 생성효소의 돌연변이 단백질들은 야생형과 같이 형광성 리간드인 6,7-dimetyl-8-ribityllumazine 혹은 리보플라빈과 결합할 수 있는 능력을 지니고 있었으나, N-RS C47D, N-RS ET66,67DQ 돌연변이 단백질의 경우는 리간드와의 결합능력이 현저히 떨어져 형광을 띠지 않았다. 대부분의 돌연변이 단백질들의 형광 세기는 야생형 단백질(N-RS wt)보다 낮았으나, N-RS C48S는 예외적으로 야생형 단백질에 비해 2배 이상의 형광세기를 가졌다. 이와 같은 결과를 바탕으로 리보플라빈 생성효소와 형광성 리간드 사이의 상호작용을 예측 할 수 있으며, N-RS C48S 돌연변이 단백질의 형광성을 활용하여 효과적으로 효소 저해제를 발굴할 수 있는 고속다중 스크리닝 법(high-throughput screening system)으로써 활용될 수 있을 것이다. Riboflavin synthase catalyzes the formation of one molecule of each riboflavin and 5-amino-6-ribitylamino-2,4-pyrimidinedione by the transfer of a 4-carbon moiety between two molecules of the substrates, 6,7-dimetyl-8-ribityllumazine. The most remarkable feature is the sequence similarity between the N-terminal half (1-97) and the C-terminal half domain (99-213). To investigate the structure and fluorescent characteristics of the N-terminal half of riboflavin synthase (N-RS) in Escherichia coli, more than 10 mutant genes coding for the mutated N-terminal domain of riboflavin synthase were generated by polymerase chain reaction. The genes coding for the proteins were inserted into pQE vector designed for easy purification of protein by 6X-His tagging system, expressed, and the proteins were purified. Almost all mutated N-terminal domain of riboflavin synthases bind to 6,7-dimethyl-8-ribityllumazine and riboflavin as fluorescent ligands. However, N-RS C47D and N-RS ET66,67DQ mutant proteins show colorless, indicating that fluorescent ligands were dissociated during purification. In addition, most mutated proteins show low fluorescent intensity comparing to N-RS wild type, whereas N-RS C48S posses stronger fluorescent intensity than that of wild type protein. Based on this result, N-RS C48S can be used as the tool for high throughput screening system for searching for the compound with inhibitory effect for the riboflavin synthase.

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