71 research outputs found

    (A) study on hospital worker's perception on the labor relations

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    병원행정학과/석사[ν•œκΈ€] μ˜λ£Œμ‘°μ§μ—μ„œμ˜ λ…Έμ‚¬λ¬Έμ œλŠ” μ΅œκ·Όμ— μ€‘μš”ν•œ 관심사가 λ˜μ—ˆλŠ”λ° μ΄λŠ” 1987λ…„ 6μ›” 29일 이후 μ •μΉ˜μ  λ³€ν™˜κΈ°λ₯Ό λ§žμ•„ λΆ„μΆœλœ μ‚°μ—…μ²΄μ˜ λ…Έλ™μš΄λ™μ΄ 병원계에도 ν™•μ‚°λ˜μ—ˆκΈ° λ•Œλ¬Έμ΄λ‹€. μ—¬λŸ¬ν˜•νƒœμ˜ μ •μΉ˜, 경제, μ‚¬νšŒ λ“± ν™˜κ²½μ˜ κΈ‰κ²©ν•œ λ³€ν™” μ†μ—μ„œ 곡읡사업체인 λ³‘μ›μ˜ 직원듀을 λŒ€μƒμœΌλ‘œ 노사관계 ν˜•νƒœμ— λŒ€ν•œ 인식, λ…Έμ‘° 및 μ‚¬μš©μžμ— λŒ€ν•œ 인식, 그리고 λ…Έμ‘° 및 μ‚¬μš©μžμ— λŒ€ν•œ 인식과 노사관계 ν˜•νƒœμ™€μ˜ 관련성을 μ•Œμ•„λ³΄κ³ μž ν•˜μ˜€λ‹€. 연ꡬλ₯Ό μœ„ν•˜μ—¬ μž„μ˜λ‘œ μ„ μ •λœ 2개의 λŒ€ν•™λ³‘μ›κ³Ό 2개의 ꡭ곡립병원 직원쀑 μ§μ’…λ³„λ‘œ λ¬΄μž‘μœ„ μΆ”μΆœν•˜μ—¬ 298λͺ…에 λŒ€ν•΄ 1990λ…„ 5μ›” 27일뢀터 5μ›” 30일 사이에 섀문쑰사λ₯Ό μ‹€μ‹œν•˜μ˜€λ‹€. κ²°κ³ΌλŠ” λ‹€μŒκ³Ό κ°™λ‹€. 1. μ‘λ‹΅μžμ˜ ν•™λ ₯, 직쒅, λ…Έμ‘°κ°€μž…μ—¬λΆ€κ°€ 노사관계 ν˜•νƒœμ— λŒ€ν•œ 인식과 ν†΅κ³„ν•™μ μœΌλ‘œ μœ μ˜ν•˜μ˜€λ‹€. 2. 노쑰에 κ°€μž…ν•œ μ‚¬λžŒμ΄ 노쑰에 κ°€μž…ν•˜μ§€ μ•Šμ€ μ‚¬λžŒλ³΄λ‹€ 노쑰에 λŒ€ν•œ 인식이 긍정적이고, μ„€λ¦½μ‹œκΈ°λ³„λ‘œ 보아 1987λ…„ 7월을 μ „ν›„ν•˜μ—¬ 과거에 λ…Έμ‘°κ°€ μ„€λ¦½λœ 병원보닀 그후에 λ…Έμ‘°κ°€ μ„€λ¦½λœ λ³‘μ›μ˜ 직원이 λ…Έμ‘°λ₯Ό κΈμ •μ μœΌλ‘œ μΈμ‹ν•˜κ³  μžˆμ—ˆλ‹€. 3. λ‹¨μ²΄ν˜‘μ•½μ— λŒ€ν•œ 인식은 μ—°λ Ή, ν•™λ ₯, κ²°ν˜Όμƒνƒœ, 직쒅, λ…Έμ‘°κ°€μž…μ—¬λΆ€μ™€ μœ μ˜ν•œ 상관관계가 μžˆμ—ˆμœΌλ©° λ…Έμ‘°κ°€ μ΅œκ·Όμ— μ„€λ¦½λœ λ³‘μ›μ—μ„œλŠ” 뢀정적인 응닡이 λ§Žμ•˜λ‹€. 4. μ‚¬μš©μžμ— λŒ€ν•œ 인식은 ν•™λ ₯, 직쒅, μ§μœ„, λ…Έμ‘°κ°€μž… 여뢀와 ν†΅κ³„ν•™μ μœΌλ‘œ μœ μ˜ν•˜μ˜€μœΌλ©°, 노쑰섀립이 1987λ…„ 이후에 이루어진 λ³‘μ›μ—μ„œλŠ” 뢀정적인 응닡이 λ§Žμ•˜λ‹€. 5. λ…Έμ‘°κ°€ 1987λ…„ 이후에 μ„€λ¦½λœ λ³‘μ›μ˜ 직원은 κ°ˆλ“±ν˜•κ³Ό 무μž₯νœ΄μ „ν˜•μ— 많이 μ‘λ‹΅ν•˜μ˜€κ³ , κ·Έ 이전에 μ„€λ¦½λœ 병원은 μ μ‘ν˜•κ³Ό ν˜‘λ™ν˜•μ— 많이 μ‘λ‹΅ν•˜μ˜€λŠ”λ° μ΄λŠ” ν†΅κ³„μ μœΌλ‘œ μœ μ˜ν•˜μ˜€λ‹€. 6. 노사관계 ν˜•νƒœλŠ” 노쑰에 λŒ€ν•œ 인식이 κΈμ •μ μΌμˆ˜λ‘ κ°ˆλ“±ν˜•μ΄λ‚˜ 무μž₯νœ΄μ „ν˜•μœΌλ‘œ, λ‹¨μ²΄ν˜‘μ•½μ— λŒ€ν•œ 인식이 κΈμ •μ μΌμˆ˜λ‘ ν˜‘λ™ν˜•μœΌλ‘œ, μ‚¬μš©μžμ— λŒ€ν•œ 인식이 κΈμ •μ μΌμˆ˜λ‘ μ μ‘ν˜•κ³Ό ν˜‘λ™ν˜•μœΌλ‘œ λ‚˜νƒ€λ‚¬λŠ”λ° μ΄λŠ” ν†΅κ³„μ μœΌλ‘œ μœ μ˜ν•˜μ˜€λ‹€. A study on hospital worker's perception on the labor relations Yun Mi Choi Graduate School of Health Science and Management, Yonsei University (Directed by Professor Kyu Sik Lee) Until July 1987, the problems of labor relations in the health service industry have not gained widespread attention. Little research has been conducted on the labor relations in the field of health industry. Most of the research on the labor relations has focused primarily on industry. The present study has a concern to identify and describe the predominant perception of hospitals workers on the labor relations. The research method used in this study was self-administered questionnaire responses. It was designed to measure the degree of agreement with each of five type of labor relations. (Conflict, Armed Truce, Power Bargaining, Accommodation. Cooperation). The pre-test was done by 20 hospital workers from May 23 to May 24, 1990. After per-test, the final questionnaire was completed. The main survey was conducted from May 27 to May 30, 1990. The Questionnaire delivered with to 370 hospital worker. A total of 315 sampled persons responed. The Response rate was 85.1% and the data were analyzed by cross tables. The results were as follows : 1. The variable with statistically significant with perception on a type of labor relation are : the educational level, type of work and whether join a union or not. 2. Those who join a union has more positively responded to the perception on a type of labor relation than one not join. 3. Perception on collective agreement was significant correlation with age, educational level, marital status, type of work and joining a union or not. 4. Perception about a manager was statistically significant with educational level, type of work, job status, joining a union or not. 5. The worker of hospital which union is organized since truly 1987, negatively responded to the perception on the manager and collective agreement. But they positively perceived on the labor union. 6. The worker of hospital which union is organized recently perceived that labor relation is "Conflict" or "Armed-Truce", while the worker of hospital which union is organized before truly 1987 perceived that labor relation is "Accommodation" or "Cooperation". [영문] Until July 1987, the problems of labor relations in the health service industry have not gained widespread attention. Little research has been conducted on the labor relations in the field of health industry. Most of the research on the labor relations has focused primarily on industry. The present study has a concern to identify and describe the predominant perception of hospitals workers on the labor relations. The research method used in this study was self-administered questionnaire responses. It was designed to measure the degree of agreement with each of five type of labor relations. (Conflict, Armed Truce, Power Bargaining, Accommodation. Cooperation). The pre-test was done by 20 hospital workers from May 23 to May 24, 1990. After per-test, the final questionnaire was completed. The main survey was conducted from May 27 to May 30, 1990. The Questionnaire delivered with to 370 hospital worker. A total of 315 sampled persons responed. The Response rate was 85.1% and the data were analyzed by cross tables. The results were as follows : 1. The variable with statistically significant with perception on a type of labor relation are : the educational level, type of work and whether join a union or not. 2. Those who join a union has more positively responded to the perception on a type of labor relation than one not join. 3. Perception on collective agreement was significant correlation with age, educational level, marital status, type of work and joining a union or not. 4. Perception about a manager was statistically significant with educational level, type of work, job status, joining a union or not. 5. The worker of hospital which union is organized since truly 1987, negatively responded to the perception on the manager and collective agreement. But they positively perceived on the labor union. 6. The worker of hospital which union is organized recently perceived that labor relation is "Conflict" or "Armed-Truce", while the worker of hospital which union is organized before truly 1987 perceived that labor relation is "Accommodation" or "Cooperation".restrictio

    Graduate Students' Perceptions on the Characteristics and Types of Good Teamwork

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    이 μ—°κ΅¬λŠ” λŒ€ν•™μ›μƒλ“€μ΄ μΈμ‹ν•˜λŠ” '쒋은 νŒ€μ›'의 νŠΉμ„±κ³Ό μœ ν˜•μ„ λΆ„μ„ν•œ ν›„, κ·Έ κ²°κ³Όλ₯Ό λ°”νƒ•μœΌλ‘œ λŒ€ν•™μ›μƒλ“€μ˜ νŒ€μ›μ—­λŸ‰ 개발 λ°©μ•ˆμ„ λͺ¨μƒ‰ν•˜λŠ”데 κ·Έ λͺ©μ μ΄ μžˆλ‹€. 이λ₯Ό μœ„ν•˜μ—¬ νŒ€μ› 및 νŒ€μ›μ—­λŸ‰μ— κ΄€λ ¨λœ 선행연ꡬλ₯Ό κ²€ν† ν•˜κ³ , λŒ€ν•™μ› 석⋅박사과정에 μž¬ν•™ 쀑인 92λͺ…μ˜ ν•™μƒλ“€μ—κ²Œ νŒ€μ„ 이루어 과업을 μˆ˜ν–‰ν•œ μžμ‹ μ˜ κ²½ν—˜μ„ 직접 μž‘μ„±ν•˜κ²Œ ν•˜λŠ” κ°œλ°©ν˜• μ§„μˆ μ„ λΆ„μ„ν•˜μ—¬ '쒋은 νŒ€μ›'의 쑰건과 μš”μΈμ„ λ²”μ£Όν™”ν•˜κ³  '쒋은 νŒ€μ›'의 μœ ν˜•μ„ 2단계 ꡰ집뢄석을 톡해 νƒμƒ‰ν•˜μ˜€λ‹€. 연ꡬ κ²°κ³Ό, 쒋은 νŒ€μ›μ€ 크게 νŒ€μ˜ μš΄μ˜λ°©μ‹, νŒ€λ¦¬λ”μ˜ νƒœλ„, νŒ€κ΅¬μ„±μ›μ˜ νƒœλ„, μ„±κ³Ό, 그리고 νŒ€κ΅¬μ„±μ›μ˜ μ—­λŸ‰ λ“±μ˜ λ‹€μ„― 가지 μš”μΈμ— μ˜ν•΄ κ²°μ •λ˜λŠ” κ²ƒμœΌλ‘œ λ°ν˜€μ‘Œκ³ , '쒋은 νŒ€μ›'의 μœ ν˜•μ—λŠ” 높은 μ„±κ³Όλ₯Ό λ‹¬μ„±ν–ˆλ˜ νŒ€μ›, νŒ€μ˜ λΆ„μœ„κΈ°λ‚˜ μ˜μ‚¬μ†Œν†΅ 및 업무뢄담 λ“± 운영 방식이 μ’‹μ•˜λ˜ νŒ€μ›, 그리고 νŒ€μ›κ³Ό νŒ€λ¦¬λ”μ˜ νƒœλ„κ°€ μ’‹μ•˜λ˜ νŒ€μ›μ˜ μ„Έ 가지가 μ‘΄μž¬ν•˜λŠ” κ²ƒμœΌλ‘œ λ°ν˜€μ‘Œλ‹€. μ΄μƒμ˜ κ²°κ³Όλ₯Ό ν† λŒ€λ‘œ ν•™λ¬Έκ³΅λ™μ²΄μ—μ„œ λŒ€ν•™μ›μƒμ˜ νŒ€μ›μ—­λŸ‰ κ°œλ°œμ„ μœ„ν•œ 쒅합적인 λ°©μ•ˆμ„ μ œμ‹œν•˜μ˜€λ‹€. 이 μ—°κ΅¬λŠ” λŒ€ν•™μ› 쑰직의 μ„±κ³Όλ₯Ό ν–₯μƒμ‹œν‚€κ³  경쟁λ ₯을 높일 수 μžˆλŠ” 핡심 μš”μ†Œλ‘œμ„œμ˜ '쒋은 νŒ€μ›'의 λͺ¨μŠ΅κ³Ό μœ ν˜•μ„ μ‚΄νŽ΄λ΄„μœΌλ‘œμ¨, λŒ€ν•™μ›μƒμ—κ²Œ μš”κ΅¬λ˜λŠ” μ—­λŸ‰μœΌλ‘œμ„œμ˜ νŒ€μ› 개발 λ°©ν–₯에 μ‹œμ‚¬μ μ„ μ œκ³΅ν•˜μ˜€λ‹€λŠ” μ μ—μ„œ μ˜μ˜κ°€ μžˆλ‹€. This study was conducted to explore the characteristics and types of good teamwork'. In order to fulfill the research questions, 92 graduate students in A University in Korea were asked to answer an open-ended questionnaire on their teamwork experiences. Based on the responses, good teamwork' was coded as five major categories: the way teams are managed, team leaders' attitudes, team members' attitudes, team performance, and team members' competencies. The five categories were divided into twelve sub-categories. They are autonomous atmosphere, effective task allocation, open communication and feedback, taking initiative, reasonable leader, shared goals, ownership, caring, high performance, psychological satisfaction, outstanding competency, and similar academic background. The types of good teamwork' were analyzed through two-step cluster analysis, which resulted in three types of good teamwork'. Based on these analyses, practical and theoretical implications for developing graduate students' teamwork competency and managing good teamwork' were suggested. Key words: teamwork, teamwork competency, teamwork development, human resource developmen

    A study on development of the managerial leadership test : a tacit knowledge approach

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    ν•™μœ„λ…Όλ¬Έ(박사) --μ„œμšΈλŒ€ν•™κ΅ λŒ€ν•™μ› :κ΅μœ‘ν•™κ³Ό,2008. 8Docto

    λ¬΄κ°€μ†Œμ œ νƒ€μž…κ³Ό λ¬΄ν”„νƒˆλ ˆμ΄νŠΈ κ°€μ†Œμ œλ₯Ό μ‚¬μš©ν•œ μΉœν™˜κ²½ PVAc 에멀젼 μ ‘μ°©μ œμ˜ λ¬Όμ„± 평가

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    Thesis(masters) --μ„œμšΈλŒ€ν•™κ΅ λŒ€ν•™μ› :μ‚°λ¦Όκ³Όν•™λΆ€(ν™˜κ²½μž¬λ£Œκ³Όν•™μ „κ³΅),2009.8.Maste

    νŒ¨μ…˜ λ””μžμΈμ˜ 창쑰적 λ°œμƒκ³Ό λͺ¨ν˜•κ°œλ°œμ— κ΄€ν•œ 연ꡬ

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    ν•™μœ„λ…Όλ¬Έ(박사)--μ„œμšΈλŒ€ν•™κ΅ λŒ€ν•™μ› :의λ₯˜ν•™κ³Ό,2001.Docto

    리 μ΄ν•΄ν•˜κΈ°οΌμ„œμ–‘μ² ν•™κ³Όμ˜ 비ꡐλ₯Ό μ€‘μ‹¬μœΌλ‘œ

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    μ–΄μ§€λŸ½κ²Œ λ³€ν•˜λŠ” 세상 μ†μ—μ„œ, λ³€ν•˜μ§€ μ•Šμ€ μ ˆλŒ€μ μΈ κ·Έ 무엇을 μ°ΎμœΌλ €λŠ” λ…Έλ ₯은 μ‹œλŒ€μ™€ λ™μ„œμ–‘μ„ λ§‰λ‘ ν•˜κ³  μΈκ°„μ˜ 주된 κ΄€μ‹¬μ‚¬μ˜€λ‹€. μ΄λŸ¬ν•œ 보편 진리 탐ꡬ에 ν˜ΈκΈ°μ‹¬μ„ 가지고 λ›°μ–΄λ“  μ‚¬λžŒλ“€μ΄ μ œμ‹œν•œ λ‹΅μ•ˆλ“€μ΄ μ–Όλ§ˆλ‚˜ λ‹€μ–‘ν•œμ§€ μš°λ¦¬λŠ” 역사 μ†μ—μ„œ 어렡지 μ•Šκ²Œ κ·Έ μ˜ˆλ“€μ„ μ°Ύμ•„λ³Ό 수 μžˆλ‹€. μ–΄λ–€ μ΄λŠ” μ΄μ„±μœΌλ‘œλ§Œ 닿을 수 μžˆλŠ” 세계에 보편 진리가 μžˆλ‹€κ³  λ§ν–ˆκ³ , 심지어 탐ꡬ κ²°κ³Ό 보편 μ§„λ¦¬λŠ” μ—†λŠ” κ²ƒμœΌ 둜 판λͺ…λ˜μ—ˆλ‹€κ³  μ£Όμž₯ν•˜λŠ” 이도 μžˆλ‹€. μ΄λ ‡κ²Œ λ³€ν™”λ¬΄μŒν•œ ν˜„μƒμ„ κΏ°λš«λŠ” κ·Έ 무엇에 λŒ€ν•œ μΈκ°„μ˜ 열망은 μ‹€λ‘œ λ‹€μ–‘ν•œ λ‹΅μ•ˆμ„ λ‚΄λ†“μ•˜μ§€λ§Œ, κ·Έ μ€‘μ—μ„œλ„ 동양철학이 μ œμ‹œν•˜λŠ” λ¦¬λŠ” μ–΄λ”˜κ°€ νŠΉλ³„ν•œ ꡬ석이 μžˆλ‹€. μ„œμ–‘μ² ν•™μ˜ 전톡과 λΉ„κ΅ν–ˆμ„ λ•Œ 리만이 κ°–λŠ” νŠΉμ§•μ„ μ€‘μ‹¬μœΌλ‘œ 리λ₯Ό μ΄ν•΄ν•˜κ³ , λΉ„νŒν•΄λ³Ό 점은 μ—†λŠ”μ§€ μ‚΄νŽ΄λ³΄λ„λ‘ ν•˜κ² λ‹€

    선별적 μ„Έμ΄ν”„κ°€λ“œμ˜ 합법성과 κ±°λŒ€ λ‹€μž μžμœ λ¬΄μ—­ν˜‘μ • μ‹œλŒ€ μ„Έμ΄ν”„κ°€λ“œ 체제의 λ°œμ „λ°©ν–₯

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    ν•™μœ„λ…Όλ¬Έ (석사)-- μ„œμšΈλŒ€ν•™κ΅ κ΅­μ œλŒ€ν•™μ› : κ΅­μ œν•™κ³Ό(κ΅­μ œμ§€μ—­ν•™μ „κ³΅), 2014. 2. μ•ˆλ•κ·Ό.With an exponential increase of FTAs since 1990s, the rule diversification problem is becoming a controversial issue. Selective safeguard measures that allow the exclusion of RTA partners from applying safeguard actions are one of those controversial issues, because they violate Article 2.2 of the Agreement on Safeguards. The WTO DSBs concept of parallelism was not enough to solve the controversies. Therefore, the first part of this thesis analyzed the three types of selective safeguard rules based on five kinds of global safeguard rules, and argued that GATT Article XXIV can justify selective safeguard measures but not all of them are legal in practice. The second part of the thesis focused on the new trend of mega-FTAs such as TPP or RCEP, where multiple countries agree to conclude a single FTA. This would lead to several systemic changes regarding selective safeguards in the FTA safeguard system. First, there will be a growing need for the sophisticated provisions for selective safeguards due to the increase in the violation of WTO rules by selective safeguards and mounting conflicts surrounding selective safeguards. Based on the analysis, the thesis suggested a desirable type of global safeguard rules in the multilateral FTAs. Second, bilateral and special safeguard measures will be increasingly important in the mega-FTAs era.TABLE OF CONTENTS ABSTRACT...............................................................i TABLE OF CONTENTS.............................................iii LIST OF TABLES AND FIGURES................................iv CHAPTER 1. Introduction...........................................1 I. Backgrounds and Motivation...................................1 II. Purpose and Structure of the Study.........................2 CHAPTER 2. WTO Safeguard System.........................3 I. Definition and Purpose of Safeguard Measures.........3 II. Non-selectivity of WTO Safeguard Measures...........5 CHAPTER 3. FTA Safeguard System and the Issue of Selective Safeguards...............................................7 I. Bilateral and Global Safeguard Measures................7 II. Various Types of Global Safeguard Rules in RTAs...7 III. Selective Safeguards in the FTA Safeguard Mechanism11 1. Definition of Selective Safeguard Measure....................11 2. Types of Global Ssafeguard Provisions Allowing Selective Safeguards...................................................12 IV. Problems of Selective Safeguard Measures...............13 CHAPTER 4. Legality of Selective Safeguards.................15 I. Positions of WTO DSB on Selective Safeguards Issue: Parallelism .................................................................15 1. Logics of Parallelism...........................................15 2. Criticism against Parallelism.....................................18 II. Legality of Selective Safeguards................................20 1. De jure Legality of Selective Safeguards................20 1.1 Permissibility of Safeguards in the FTA System......21 1.2 Justification of the Violation of the Agreement on Safeguards by GATT Article XXIV ................................23 2. De facto Legality of Each Type of Selective Safeguard Measures in Practice..................................................28 CHAPTER 5. The Future of Selective Safeguards in the mega-FTAs Era.........................................................31 I. The New Trend of Regionalism: Mega-FTAs..............31 II.The Future of Selective Safeguards in the Mega-FTAs Era .........................................................................33 1. Need for the Sophisticated Provisions for Selective Safeguards.................................................................33 1.1 Increase in the Violation of WTO rules by Selective Safeguards...........................................33 1.2 Mounting Conflicts Surrounding Selective Safeguards.................................................................35 1.3 Desirable provisions for selective safeguards..........36 2. Increasing Importance of Bilateral and Special Safeguards .........................................................................39 CHAPTER 6. Conclusion..............................................42 APPENDIX.................................................................44 BIBLIOGRAPHY..........................................................45 ꡭ문초둝....................................................................47Maste

    (A)Reconsideration over adaptation effects with a restored phoneme : based on scoustic signals

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    Thesis(master`s)--μ„œμšΈλŒ€ν•™κ΅ λŒ€ν•™μ› :심리학과 지각심리학전곡,2004.Maste

    Observation of Lipid Mixing and Content Release for SNARE-mediated Intracellular Membrane Fusion by ALEX-FRET

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    ν•™μœ„λ…Όλ¬Έ (석사)-- μ„œμšΈλŒ€ν•™κ΅ λŒ€ν•™μ› : ν™”ν•™λΆ€, 2012. 2. κΉ€μ„±κ·Ό.λ³Έ μ—°κ΅¬λŠ” 생λͺ… ν˜„μƒμ˜ 기본이 λ˜λŠ” 세포막 μœ΅ν•©κ³Ό 이에 λ”°λ₯Έ μ‹ κ²½ 전달 물질의 전달이 μ–΄λ– ν•œ λ°©μ‹μœΌλ‘œ μΌμ–΄λ‚˜λŠ”μ§€λ₯Ό μ‚΄νŽ΄λ³΄κ³  μžˆλ‹€. μ‹ κ²½ 전달 λ¬Όμ§ˆμ€ μ‹œλƒ…μŠ€μ—μ„œ μ†Œν¬κ°€ ν˜•μ„±λ˜λ©΄μ„œ κ·Έ μ•ˆμ— ν¬ν•¨λ˜κ³  세포막 μœ΅ν•© 과정을 톡해 λ‹€λ₯Έ μ„Έν¬λ‘œ 전달될 수 μžˆλŠ”λ° μ΄λŸ¬ν•œ 일련의 과정을 λ‹¨λΆ„μž μˆ˜μ€€μ—μ„œ 규λͺ…ν•˜κ³ μž ν•œλ‹€. 특히, Soluble NSF Attachment Protein Receptor(SNARE)은 세포막에 μ‘΄μž¬ν•˜λŠ” λ‹¨λ°±μ§ˆμ˜ 총칭으둜 λ‰΄λŸ°μ—μ„œ μ†Œν¬λ‘œ λ‘˜λŸ¬μ‹ΈμΈ μ‹ κ²½μ „λ‹¬λ¬Όμ§ˆμ΄ λ‹€μŒ λ‰΄λŸ°μ΄λ‚˜ μ„Έν¬λ‘œ μ „λ‹¬λ˜λŠ” 세포막 μœ΅ν•©κ³Όμ •μ— κ΄€μ—¬ν•œλ‹€. μ΄μ „μ˜ μ—°κ΅¬λŠ” μ „κΈ°μ˜λ™λ²•κ³Ό 같이 μ•™μƒλΈ”μ—μ„œ μ§„ν–‰λ˜μ–΄ μ™”κΈ° λ•Œλ¬Έμ— 평균화에 μ˜ν•œ 였λ₯˜λ₯Ό ν”Όν•  수 μ—†μœΌλ©°, μ‹œκ°„μ  동기화 과정이 μΆ”κ°€μ μœΌλ‘œ ν•„μš”ν•˜λ‹€. μ΄λŸ¬ν•œ ν•œκ³„μ μ„ κ·Ήλ³΅ν•˜κΈ° μœ„ν•΄ 기쑴에 κ΄€μ°°ν•  수 μ—†μ—ˆλ˜ λΆ„μž ν•˜λ‚˜μ˜ μ‹€μ‹œκ°„ κ΅¬μ‘°λ³€ν™”λ‚˜ μ›€μ§μž„, 속도둠적 뢄석이 κ°€λŠ₯ν•œ λ‹¨λΆ„μž λΆ„κ΄‘ν•™ 기술이 λ°œμ „ν•΄ μ™”λ‹€. 그리고 λΆ„μžκ°„μ˜ 거리λ₯Ό λ‚˜λ…Έλ―Έν„° μˆ˜μ€€μ—μ„œ μ•Œ 수 μžˆλŠ” FRETν˜„μƒμ΄ λ„μž…λ˜μ—ˆλ‹€. 특히 λ³Έ 연ꡬ에 μ‚¬μš©λœALEX-FRET의 기술 개발둜 λ™μ‹œλ‹€λ°œμ μΈ ν˜„μƒμ„ κ΄€μΈ‘ν•  수 있게 λ˜μ—ˆλ‹€. μ΄λŸ¬ν•œ ALEX-FRET을 μ΄μš©ν•˜μ—¬ in vitroμ—μ„œ λͺ¨λΈ μ‹œμŠ€ν…œμœΌλ‘œ ν•©μ„±ν•œ vesicle을 기반으둜 생체 λ‚΄μ—μ„œ 유기적으둜 μΌμ–΄λ‚˜λŠ” μ‹ κ²½μ „λ‹¬μ˜ 일련의 과정을 ν™•μΈν•˜κ³ μž ν•˜μ˜€λ‹€. λ³Έ μ—°κ΅¬μ—μ„œλŠ” κ΅μ°¨λ ˆμ΄μ € κΈ°μˆ μ„ λ„μž…ν•œ λΆ„κ΄‘ν•™ κΈ°μˆ μ„ μ ‘λͺ©ν•˜μ—¬ SNARE λ‹¨λ°±μ§ˆμ„ 맀개체둜 세포막 μœ΅ν•©κ³Ό μ‹ κ²½ 전달 물질의 전달 과정을 λ‹¨λΆ„μž μˆ˜μ€€μ—μ„œ λ™μ‹œ κ΄€μΈ‘ν•¨μœΌλ‘œμ¨ 이에 λŒ€ν•œ λ©”μ»€λ‹ˆμ¦˜μ„ λ°νžˆλŠ” 것을 μ΅œμ’… λͺ©ν‘œλ‘œ ν•˜μ˜€λ‹€.The goal of this study is to observe how membranes are fused and the neurotransmitters are released in chemical synapse at single vesicle level. Especially, it is well known that soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) play an important role to accelerate intracellular membrane fusion. That is, we tried to demonstrate consecutive events of membrane fusion using model system, proteoliposomes reconstituted with SNARE proteins in vitro. The previous study is limited to ensemble assay such as electrophoresis whose problems are ensemble averaging and synchronization process. To overcome these limitations, many techniques are developed for detecting single molecule. Among several single molecule fluorescence spectroscopy techniques, alternating-laser excitation (ALEX) is the unique tool to observe complicate events simultaneously and thats why we used ALEX in this study. First of all, fluorescence resonance energy transfer (FRET) has an effect on development of single molecule fluorescence spectroscopy by introducing a second fluorophore. FRET is a mechanism describing energy transfer from a donor fluorophore in its electronic excited state to an acceptor fluorophore through non-radiative dipole-dipole coupling. Since FRET efficiency depends sensitively on the donor to acceptor distance within 10nm, it is very powerful to observe bio-molecules in regard to comparable scale. In this study, we used ALEX which is a confocal microscopy combining alternating laser. By using alternating laser system, we can define new value, stoichiometry (S) which is able to analyze the combination of fluorescence labeling on target molecules. In other words, ALEX-FRET can measure not only the stoichiometry values but also information for the distance between two fluorophores. As a model system for intracellular membrane fusion, we employed SNARE-containing vesicles labeled with DiO and DiI and carried out FRET experiments for lipid mixing as well as content release at the single vesicle level. By utilizing the capability of 3-color ALEX-FRET, we tried to demonstrate the content mixing of vesicles when Cy5/quencher dual-labeled molecular beacon encapsulated in a vesicle encountered its complementary DNA in the other vesicle. This study for underlying whole process of biology would be solution to solve the previous limitation, and milestone leading the way of research in biotechnology.Maste
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