Serratia marcescens에서 cAMP receptor protein(CRP)유전자의 클로닝 및 해석

유주순,문종환,정수열,김혜선,최용락 東亞大學校附設遺傳工學硏究所 1999 遺傳工學硏究 Vol.- No.6

전사조절인자로서 잘 알려져 있는 cAMP receptor protein(CRP)은 cAMP와 DNA에 결합하는 특별한 활성을 가지고 있으며, cAMP-CRP complex를 형성하여 수많은 유전자의 발현조절에 관여한다. 이러한 측면에서 cAMP-CRP의 조절은 어떤 면에서 총체적 조절체계라고까지 한다. 본 연구는 Serratia 균주에서 crp유전자의 분자적 특성 및 cAMP에 의한 발현조절을 받는 분자기구를 해석하고자 유전자를 클로닝하고 발현을 확인하였다. MacConkey배지에서 maltose를 탄소원으로 충분히 이용하지 못하는 대장균 TP2139(△crp,△lac)를 숙주로 이용하고, 염색체DNA를 library로 작성하여 얻은 형질전환체 약 일만개의 콜로니에서 red colony를 나타내는 5종류 의 양성 클론을 얻었다. 이들 클론을 Southern방법으로 확인한 결과 3kb의 단편을 가진 pCKB12클론이 crp유전자를 coding하고 있음을 확인하였다. glpD-lacZ 융합 plasmid인 pLDC6 의 BamHI부위에 pCKB12의 3kb 단편을 삽입시킨 재조합 plasmid pLDC 6-Scrp를 작성하여, 클로닝된 Serratia 의 crp 유전자가 대장균에서 유전자 전사조절에 미치는 영향을 확인한 결과 cAMP-CRP 복합체 형성에 의한 전사조절 기능이 확인되어졌다. One of the better-characterized transcription factor of E. coli is the cAMP receptor protein(CRP) and the CRP binds cAMP and DNA. The cAMP-CRP complex is involved in regulation of many genes at bacteria. The cAMP-CRP regulatory element represents, in some respects, a global regulatory network. The aim of this work was to study the structure and the mechanisms controlling the expression of CRP in Serratia marcescens. We have been get 5 different clones from Serratia which stimulated the cells to use maltose as a sole carbon source in E. coli TP2139. The crp gene clone, pCKB12, was confirmed by Southern hybridization with E. coli crp gene. The location of the crp gene was determined by constructing subclones carrying various portions of pCKB12. To investigate the potential role of CRP in E. cloi, lacZ fused plasmids were constructed and investigated the β-galactosidase activity of the fused plasmid. The Serratia marcescens cAMP receptor protein can substitute the E. coli CRP in transcriptional activation at the lacZ gene. These results suggest that Serratia marcescens cAMP receptor protein complex functions to regulate several promoters in E. coli.

PLZT 세라믹의 상변이 특성에 관한 연구

유주현,홍순석,박창엽 연세대학교 산업기술연구소 1986 논문집 Vol.18 No.2

PLZT electrooptic ceramics of compositions in the range X/65/35 with 5?? X ??9, where X is the mol % La substixuted for lead in a ceramic with a Zr/Ti ratio of 65/35, were fabricated by the two stage sintering technique. For the ceramics, dielectric properties and phase transition were investigated. X-ray diffraction analysis was performed to analize α-β phase transition. For X=5,6, its crystal structure was Rhombohedral in the thermally depoled state as well as the poled state. For 6〈X〈9, phase transition was occured by poling and the intermediate region existed between the paraelectric and ferroelectric states. For X=9, its crystal structure was the same cubic phase in the thermally depoled state and the poled state.

일부여자대학생의 Cellulite 증상의 정도와 관련요인

이순혜,류규수,김문주 慶山大學校 2000 論文集 Vol.18 No.1

A study on the degree of cellulite and the factors affecting cellulite was conducted to understand the occurrence frequency of cellulite and the degree of cellulite, for students who are belonged to department of beauty artist in junior college. 240(50.3%) out of 477 had the standard shape of body, 99(20.8%) had the big frame and heavy weight, 75(15.7%) had the typical woman shape of body, and 63(13.2%) were tall and slim. 204(42.8%) out of 477 had the dry skin,186(39.0%) had the fatty skin and 87(18.2%) had the normal skin. The results of the study are summarized as follows ; 1. The cellulite condition, the physiological factor, the psychological factor(p<0.001), the diet factor, the constitutional factor(p<0.01) and the physiological factor (p<0.05) were different, by school year. The cellulite condition was different by the place which school was in(p<0.001).By shape of body, the cellulite condition, the habit of life, the psychological factor, the physiological factor (P<0.001) and the diet factor, the constitutional factor(P<0.05) were different. The cellulite condition, the physiological factor(p<0.01) and the constitutional factor(p<0.001) were different by degree of corpulence. The habit of life(p<0.01), the constitutional factor (p<0.05) were different by religion, and the diet factor was different by status of economical class (p<0.05). The diet factor (p<0.001), the cellulite condition, the psychological factor (p<0.01)and the physiological factor, the physiological factor (p<0.05) were different by status of skin. The physiological factor (P<0.001), the cellulite condition, the habit of life, the diet factor, the constitutional factor(p<0.01) and the psychological factor (p<0.05) were different by place which student lived in. 2. Each factor has positive relation with cellulite condition. The correlation coefficients of factors were the physiological factor (0.565), the diet factor (0,433),the habit of life (0.428), the constitutional factor (0.271) and the psychological factor (0.264) respectively. 3. According to the result of regression analysis between each factor and cellulite condition, the regression coefficient of the diet factor was 0.236, that of the habit of life was 0.206, that of the constitutional factor was 0.086, and that of the psychological factor the constitutional factor was 0.108 (P<.001).

자아존중감, 가족의 건강성, 사회인구학적 변인이 농촌청소년의 학교생활적응에 미치는 영향

양순미,유영주 대한가정학회 2003 Human Ecology Research(HER) Vol.41 No.9

Thii study examined the degree of school life adjustment in d adolescents as well as related variables. For thii purpose, the data of 192 persons from 40 villages in 8 provinces were analyzed using a SAS 8.1 program. The results of the analysis were as follows : 1) The overall degree of school life adjustment was quantified as about 67 points out of a 100 possible points. 2) The variables that affected the level of adjustment were self-esteem, healthiness of family, age. 3) The explanatoy power of such variables as sex, religion, personality, school grades, father satisfaction,

다량의 하부위장관출혈이 병발된 Vibrio vulnificus 패혈증 1예

최주연,유진홍,한석원,이대훈,최민호,박순민,김연식,김선우,신완식,강문원,강성구 대한화학요법학회 1996 대한화학요법학회지 Vol.14 No.2

최근 저자들은 Vibrio vulnificus 감염에 의해 패혈증 및 이차성 피부 병변을 보인 42세의 남자 환자의 치료중 하부 위장관 출혈이 병발된 1예를 경험하였기에 보고하는 바이다. We experienced a case of Vibrop vulnificus septicemia which showed an unusual manifestation such as hematochezia. The patient was a 42-year-old man with chronic liver disease. The blood and wound culture revealed the organism. During intensive care, he suffered from massive hematochezi for several days. Colonoscopy and abdominal CT showed findings suspicious of ischemic colitis. To our knowledge, this is the first unusual case of V.vulnificus infection which showed massive lower gastrointestinal bleeding.

Flavobacterium meningosepticum에 의한 균혈증 1예

장성원,유진홍,진승원,김선우,김연식,박순민,이대훈,최민호,최주연,신완식,강문원,강지민 대한화학요법학회 1997 대한화학요법학회지 Vol.15 No.1

1996년 6월 카톨릭대학교 부천성가병원에 뇌경색으로 입원했던 61세 남자 환자의 혈액에서 Flavobacterium meningosepticum을 분리하였다. 환자는 폐렴을 동반한 패혈증의 예로 ceftriaxone과 metronidazole을 투여하였으나 입원 48시간만에 사망하였다. Flabovacterium meningosepticum is a rae cause of nosocomial infection which shows multi-drug resistance. It mainly invades patients with impaired immunity. Recently, we experienced a case of F. meningosepticum bacteremia in a patient with chronic debilitated state owing to stroke. The clinical progress was so rapid that he died within 48 hours. Blood culture revealed F. meningosepticum which showed characteristic yellow colony in blood agar plate.

Cellulose 생합성균주 Acetobacter sp. CBⅡ의 분리 및 특성

이승진,유주순,최용락 동아대학교 환경문제연구소 2000 硏究報告 Vol.23 No.1

The strain of cellulose-producing CBⅡ was isolated from the traditionally fermented persimmon vinegar, and identified as Acetobacter sp. CBⅡ with respect to physiological and biochemical characteristics. The isolated strain was cultivated statically in broth culture, a thick cellulose pellicle was formed. It was found that the strain was fluorecenced with UV light when the strain was cultivated agar medium contained Tinopal. The strain produced acetic acid from ethanol, and then decomposed acetate to CO₂ and H₂O. Capacity of cellulose biosynthesis from isolated strain was compared with Acetobacter xylinum ATCC 53582. The cellulose produced by the isolated strain was purified and cegraded by HCl. The released sugar was separated on thin layer chromatograpy(TLC) consistent with the degradation product of native cellulose.

Escherichia coli 패혈증 환자에 합병된 대칭적 하지 말단 괴사증 1예

남해성,유진홍,권순석,민준기,조현선,박민경,심병주,남유정,이지인,김진수,길욱현,조근종,신완식 대한감염학회 2005 감염과 화학요법 Vol.37 No.6

We have encountered a rare case of symmetrical peripheral gangrene complicating Escherichia coli sepsis in a 47-years-old male. He was successfully treated with antibiotics, anticoagulants, and vasodilator. To our knowledge, this is the first report on symmetrical peripheral gangrene complicating E. coli sepsis in Korea.

Cellulose 생합성 세균의 분리 및 특성

이승진,유주순,정수열,최용락 東亞大學校附設遺傳工學硏究所 1998 遺傳工學硏究 Vol.- No.5

발효시켜서 만드는 감식초에서 시료를 채취하여 배양하고, pellicle을 형성하는 single colony를 cellulose생합성균으로 분리하였다. 분리된 균주는 형태적 특성, 알콜의 재산화 등의 생리, 생화학적 특성 등에 의하여 Acetobacter속으로 분류되었으며, Acetobacter CBI-2라고 명명하였다. Acetobacter CBI-2는 정지배양 시에 기존의 생합성 균주로 알려진 Acetobacter xylinum과 대등한 cellulose생합성 능력을 나타내었다. Acetobacter CBI-2이 생성한 고분자물질의 분해산물을 TLC법으로 확인한 결과 기존 cellulose의 것과 일치하였다. Acetobacter CBI-2에서 genomic DNA를 분리 정제하여 cel A 영역을 probe로 하여 hybridization시킨 결과, 상동성을 나타내어 분리 균주에는 cellulose 생합성 관련 유전자가 존재함을 나타내었다.(1997년 1월 7일 접수, 1997년 2월 24일 수리). A screening was performed to isolate the cellulose-producing microorganisms from vinegar in Korea. The isolated strain was identified as Acetobacter sp. with respect to physiological and biochemical characteristics and designated as Acetobacter CBI-2. Cellulose production of Acetobacter CBI-2 was equal with the well known cellulose-producing bacteria, A.xylinum. The result of separation on thin layer chromatography (TLC) was consistent with the degradation product of native cellulose. The presence of genes required for the cellulose biosynthesis in Acetobacter CBI-2 was confirmed by Southern hybrization.

대장균 sfs 유전자의 전사조절

이희영,유주순,정수열,이승진,최용락 東亞大學校附設遺傳工學硏究所 1996 遺傳工學硏究 Vol.- No.3

sfs(sugar fermentation stimulation) 유전자는 crp*¹이 도입된 MK2001 균주에서 mal 유전자의 발현을 촉진시키는 특성으로서 클로닝 되어졌다. sfs4 유전자의 발현은 두 개의 promoter에 의하여 조절되어진다. 본 연구는 이들 두 promoter (P₁과 P₂)의 cAMP-CRP에 의한 발현조절을 보고자 조절영역과 CRP 결합영역에 변이가 도입된 DNA가 in vitro transcription 방법에 의해 확인되어졌다. 두 개의 promoter가 공존시에는 cAMP에 의한 P₂의 전사는 촉진되어졌으며, P₁의 전사는 억제되어졌다. P₂만이 존재할 때에도 전사는 cAMP에 의하여 촉진되어졌다. CRP 결합부위에서의 변이는 cAMP에 의한 전사촉진이 크게 감소하였다. sfs7 (nlp) 유전자의 전사산물을 확인하였으며, 이는 cAMP에 의한 발현조절을 받지 않았다. sfs(sugar fermentation stimulation) genes are involved in regulation of mal gene expression with the altered crp*¹ gene. Expression of the sfs4 gene appeared to be regulated by a two promoters. In this research, the cAMP-CRP dependent regulation of P₁ and P₂ promoter of sfs4 was examined by in vitro transcription using the regulatory region and mutated DNA of the CRP binding region. The transcription of sfs4 from P₂ is stimulated, but P₁ was totally repressed by in vitro transcription. In mutant in the CRP binding region, the effect of activation by cAMP was severely reduced. The transcript of sfs7(nlp) was identified and regulation by cAMP was not observed at the mRNA level.