비만 관련인자와 ALT 증가와의 관련성

민홍기,이정규,이하린,박현지,김윤진,이상엽 부산대학교 병원 암연구소 2009 부산대병원학술지 Vol.- No.25

Antioxidant Activity and Whitening Effect of Lindera erythrocarpa Extract

Min ?Chul Kang,Ryeo Kyeoung Ko,Ju-Yeop Lee,Haeng-Bum Kim,Bong-Seok Kim,Jong-Heon Han,Seong-Hyun An,Gi-Ok Kim 한국생물공학회 2008 한국생물공학회 학술대회 Vol.2008 No.10

플라이애시를 다량 사용한 콘크리트의 수화발열 특성과 압축강도 특성에 관한 실험적 연구

조규현,박무영,백민수,김우재,임남기,정상진 대한건축학회 2003 대한건축학회 학술발표대회 논문집 - 계획계/구조계 Vol.23 No.2

This study is for the great quantity use of fly-ash. For the producing of high volume concrete from the use of fly-ash, the method of replacement between bonding agents and fine aggregate by fly-ash at the slt was used that the adiabatic temperature rise of concrete about the mass member which bad been produced by the method that was mentioned before, and the hydration heat of the core test pieces in concrete was measured. Also the core test pieces which were replaced with fly-ash was studied by the compressive strength's comparison between standard care test pieces and core test pieces. In the case of mass test pieces, hydration heat and the time to reach the highest temperature were decreased by an increase in replaced fly-ash's amounts of concrete. In addition, among the test pieces having the same amounts of concrete, the test pieces having more replaced amounts of fly-ash's fine aggregate showed higher hydration heat and the increased time to reach the highest temperature. Compressive strength was also increased by hydration heat's decrease according to fly-ash replacement. Replacement of fly-ash was more effective in high temperature environment.

Clozapine이 백서의 Schedule-Induced Polydipsia에 미치는 영향

이기철,정홍경,이정호,홍승범,최영민,전성일,정재현,하준명 대한생물치료정신의학회 2000 생물치료정신의학 Vol.6 No.2

Object : This study was designed to evaluate the effects of clozapine which is one of most useful atypical antipsychotics in the schedule-induced polydipsic rat which is an animal model of obsessive-compulsive disorder. Methods : Spraque-Dawley rats were placed in automatic cage where a pellet dispenser automatically dispensed 90mg pellets on a fixed-time 60 seconds(FT- 60s) feeding schedule over 150-minute test session for 4 weeks. After 4 weeks of daily exposure to the FT-60s feeding schedule, experimental rats met a predetermined criterion for polydipsic behavior(greater than 3 times of water per session on average). Rats were stratified into clozapine(0.34mg/kg,i.p.), clozapine(14.63mg/kg,i.p.), clomlpramine(5mg/kg,i .p.), and vehicle (1cc/kg,i.p.) group and treated with each drugs for 3 weeks. To identify the non-polydipsic food-deprived rats, a separate group of rats(N=8) were individually housed and given a single bolus(14.5gm) of food per day which maintained them at their average body weight. Results : The results were as follows ; 1)After 4 weeks of scheduled feeding procedure, the experimental group showed significant differences than bolus control in the amount of water consumption as compared with their baseline of water intakes for 4 weeks. At the same periods, there were no differences between the experimental group and bolus control in the borty weight. 2)The clomipramine group, the clozapine 0.34mg group and the clozapine 14.63mg group showed significant decrease in the amount of water intake for at 2nd & 3rd week of drug treatment as compared with their baseline of polydipsic water intakes. But, the vehicle control group showed no changes of amounts of water intake for 3 weeks of treatment as compared with their baseline of polydipsic water intakes. Conclusion : Above findings suggest that the fixed time feeding Procedure for Schedule induced polydipsia as an animal model of obsessive compulsive disorder was effective to the evaluation of pharmacological challenge study. In clinical situation, the authors suggest that atypical antipsychotic drugs which act as serotonin and dopamine receptor antagonist may be helpful to improve the symptoms of the patients with treatment refractory obsessive-compulsive disorder.

세로토닌성 항우울제가 백서의 Schedule-Induced Polydipsia에 미치는 영향

이기철,이정호,박중섭,최영민,전성일,정홍경,하준명,정재현 대한생물치료정신의학회 1999 생물치료정신의학 Vol.5 No.2

Object : Schedule-induced polydipsia is considered as an animal model of obsessive-compulsive disorder inrats. The authors evaluated the chronic effects of fluoxetine and clomipramine as serotonergic antidepressants and haloperidol as dopaminergic antagonist on the schedule-induced polydipsia in rat.Methods : Spraque-Dawley rats weighing 200-250gm were individually housed, maintained and allowed free access to water for 1 week. And then the rats were placed on a restricted diet. To induce polydipsia, rats were placed in automatic cage where a pellet dispenser automatically dispensed 90mg pellets on a fixed-time 60 seconds(FT 60s) feeding schedule over 150-minute test session for a day. Water was available at all times during the feeding schedule in automatic cage. After 4 weeks of daily exposure to the FT 60s feeding schedule, experimental rats met a predetermined criterion for polydipsic behavior(greater than 3 times of water per session on average). 4 groups of rats were administered fluoxetine(5mg/kg/i.p.), clomipramine(5mg/kg/i.p.), haloperidol(0.1mg/kg/i.p.), vehicle(1cc/kg/i.p.) for 3 weeks. Rats were tested once a week to access schedule induced polydipsic behavior. The chronic effects of experimental drugs on schedule induced polydipsic behavior were analyzed with repeated analysis of variance and Scheffe test as a post-hoc comparison.In order to measure water consumption in non-polydipsic food-deprived rats, a separate group of rats(N=8) were individually housed and given a single bolus(14.5 gm) of food per day which maintained them at their average body weight.Results and Conclusion : The results were as follows ;1) After 4 weeks of daily feeding procedure with fixed time schedule for 60 seconds per day, the experimental group showed significant differences than the control in the amount of water consumption as compared with their baseline water intakes. At the same periods, there were no differences between the experimental group and the control in body weight. 2) The clomipramine treated group and the fluoxetine treated group showed significant decrease in the amount of water intake as compared with their baseline of polydipsic water intakes for 3 weeks of treatment. However, the haloperidol treated group and the vehicle control group showed no changes of amounts of water intake for 3 weeks of treatment as compared with their baseline of polydipsic water intakes. 3) At 2 weeks of drug treatment, clmipramine treated group(16.88±6.51ml) and the fluoxetine treated group(22.50±10.35ml) showed significantly lower amounts of water intake than the haloperidol treated group (41.25±7.06ml) or vehicle control group(37.50±12.54ml). And also the clomipramine treated group(13.75±5.18ml) and the fluoxetine treated group(18.75±3.54ml) showed significantly lower amounts of water intake than the haloperidol group(35.00±11.65ml) and the vehicle control(34.38±6.78ml) at 3 weeks of drug treatment. Above findings suggest that the fixed time feeding procedure for schedule-induced polydipsia as an animal model of obsessive compulsive disorder was effective to the evaluation of pharmacological challenge study. The author confirmed that schedule-induced polydipsia was successfully decreased for 3 weeks of administration of clomipramine and fluoxetine but there was no response to haloperidol.

Risperidone이 백서의 Schedule-Induced Polydipsia에 미치는 영향

이기철,이정호,윤도준,최영민,전성일,김태수,정홍경,하준명,정재현 大韓神經精神醫學會 2000 신경정신의학 Vol.39 No.3

연구목적 : 강박장애의 원인론중 세로토닌-도파민 가설에 기초하여 강박장애 동물모형으로 고려되는 고정된 시간 간격으로 평소의 먹이섭취량보다 작은 양의 음식물을 백서에게 장기간 공급하여 다음중(schedule-induced polydipsia : SIP)을 유발시켰다. SIP모형에 강박장애에 효과적이라고 알려진 선택적 세로토닌 재흡수 차단제로서 fluoxetine을 장기 투여하고, 신경절후 5-HT²와 도파민 D² 수용체를 동시에 차단하는 risperidone을 투여하고, 신경절후 도파민 수용체 차단제인 haloperidol을 투여하였다. 그결과로서 risperidone이 강박장애 동물모형으로 고려되는 SIP에서 어떤 영향을 미치는지 알아보았다. 방 법 : SIP를 유발하기 위해 각각의 사육상자에 1정당 90㎎의 사료를 자동급이장치 (automatic dispenser)에서 60초당 1정씩 고정된 시간 간격으로 하루에 150분씩 공급하였다. 4주간 고정된 시간 간격으로 머기를 공급하고 매주 음수량과 체중을 측정한 실험 동물과 동등한 사료의 양을 한번에 덩어리로 공급받은 통제 집단의 체중과 음수량을 비교하였다. SIP 행동변화를 보인 실험 동물들을 fluoxetine 5㎎/㎏(N=8), risperidone 0.1㎎/㎏(N=8), haloperidol 0.1㎎/㎏(N=8), 그리고 vehicle 대조군 1㏄/㎏(N=8)으로 나누고 각각의 실험 동물군에서 3주간에 걸쳐서 실험 약물을 매일 복강내 주사하였다. 매주 실험 동물의 음수량과 체중을 측정, 비교하였다. 결 과 : 1) 고정된 시간 간격으로 제한된 먹이를 공급한 실험 동물군은 1주부터 4주에 걸쳐서 기저치보다 유의한 음수량의 증가를 보였다. 반면 통제 집단은 2주째 음수량이 일시적으로 증가한 소견 이외에 4주간의 실험 기간중 유의한 변화는 보이지 않았다. 실험 동물과 통제 집단간의 음수량에서 3주와 4주째에 실험 동물이 통제 집단보다 유의하게 높은 음수량을 보였지만 양군간에 체중의 차이는 보이지 않았다. 2) 각각의 실험 동물군 내에서 risperidone 0.1㎎ 투여군은 약물 투여 2주부터 3주까지 기저치 음수량과 비교하여 유의한 저하를 보였다. Risperidone 0.5㎎ 투여군은 약물터여 3주에서 기저치의 음수량과 비교하여 유의한 저하를 보였다. Fluoxetine 투여군은 약물 투여 시작 1주부터 3주에서 기저치의 음수량과 비교하여 유의한 저하를 보였다. 한편, haloperidol 투여군과, vehicle은 3주간에 걸친 약물 투여에서 각각의 기저치 음수량과 비교하여 차이를 보이지 않았다. 3) 실험 동물 각 군간에 약물 투여 시간 경과에 따른 음수량을 비교한 바, 약물투여 1주에서 각 군간에 유의한 차이는 없었다. 약물 토여 2주에서 fluoxetine 투여군, risperidone 0.1㎎ 투여군, 그리고 risperidone 0.5㎎투여군이 haloperidol 투여군과 비교하여 유의한 차이를 보였다. 약물투여 3주째에 fluoxetine투여군, risperidone 0.1㎎ 투여군, 그리고 risperidone 0.5㎎투여군이 haloperidol 투여군, vehicle과 비교하여 유의한 음수량의 저하를 보였다. 결 론 : 백서의 강박 행동은 fluoxetine, risperidone에 의해 효과적으로 억제되었으나 haloperidol에는 반응이 없었으므로, 임상에서 난치성 강박장애의 치료에 비정형 항정신병 약물 투여를 고려해 볼 수 있다고 제안한다. Objectives : This study was designed to evaluate the effects of risperidone on the schedule-induced polydipsia(SIP) which is one of animal model of obsessive-compulsive disorder in rats. We administered risperidone as a serotonin and dopamine blocking agent, fluoxetine as a selective serotonin reuptake inhibitor, and haloperidol as a dopamine antagonist to rats which showed schedule-induced polydipsic behaviour. Methods : Sprage-Dawley rats weighing 200∼250gm were individually housed and maintained and allowed free access to water. The rats were placed on a restricted diet. To induce polydipsia, rats were placed in the cage where a pellet dispenser automatically dispensed 90㎎ pellets on a fixed-time 60 seconds(FT 60s) feeding schedule over 150 minute test session per day. Water was available at all times in the cage. After 4 weeks of daily exposure to the FT 60s feeding schedule, experimental rats met a predetermined criterion for polydipsic behavior(greater than 3 times of water per session on average). 5 groups of rats were administered risperidone(0.1㎎/㎏, i.p), risperidone(0.5㎎/㎏, i.p), fluoxetine(5㎎/㎏, i.p), haloperidol(0.1㎎/㎏, i.p), and vehicle(1㏄/㎏, i.p) for 3 weeks. The rats were tested once a week to access schedule induced polydipsic behavior. Water bottles were weighed before and after the 150-minute test session. The chronic effects of administration of experimental drugs on schedule induced polydipsic behavior were analyzed with ANOVA and Scheffe test as a post-hoc comparison. In order to measure water consumption in non-polydipsic food-deprived rats, a separate group of rats(N=8) was individually housed and given a single bolus(14.5gm) of food per day which maintained them at their average body weight. Results : The results were as follows ; 1) After 4 weeks of scheduled feeding procedure, the experimental group showed significant differences than the bolus control in the amount of water consumption as compared with their average water intakes for 4 weeks. At the same periods, there were no differences between the experimental group and the bolus control in the body weight. 2) The fluoxetine group showed significant decrease in the amount of water intake at 1st, 2nd, and 3rd weeks of drug treatment as compared with their average amount of polydipsic water intakes. The risperidone 0.1㎎ group and the risperidone 0.5㎎ group showed significant decrease in the amount of water intake at the 3rd weeks of drug treatment as compared with their baseline of polydipsic water intakes. However, the haloperidol group and the vehicle control group showed no changes of amounts of water intake for 3 weeks of treatment as compared with their baseline of polydipsic water intakes. 3) The fluoxetine group(22.5±10.4ml) showed significantly lower amounts of water intake than haloperidol group(41.3±7.1ml) at 2nd weeks of drug treatment. And also the fluoxetine group(18.8±3.5ml) showed significantly lower amounts of water intake than the haloperidol group(35.0±11.7ml) and the vehicle control(34.4±6.8ml) at 3rd weeks of drug treatment. The risperidone 0.1㎎ group and the risperidone 0.5㎎ group showed significantly lower amounts of water intake than the haloperidol group(35±11.7ml) at 2nd weeks and the vehicle control(37.5±12.5, 34.4±6.8ml) at 2nd and 3rd weeks of drug treatment. Conclusions : Above findings suggest that the fixed time feeding procedure for schedule induced polydipsia could be applied as an effective animal model of obsessive compulsive disorder for the evaluation of pharmacological challenge study. We confirmed that chronic treatment with risperidone revealed antipolydipsic effect as effective as fluoxetine on the schedule-induced polydipsic behaviour but the onset of effect was later than fluoxetine.

정신분열병 및 정신분열형 장애에 대한 올란자핀의 효과와 안전성

안용민,강대엽,권준수,김창윤,김철응,반건호,신영민,이기철,이동우,이중서,조현상,채정호,김용식 大韓神經精神醫學會 2001 신경정신의학 Vol.40 No.4

연구목적 : 본 다기관 공동임상연구는 국내 환자를 대상으로 새로운 항정신병약물인 올라자핀의 치료 효과와 안전성을 확인하고자 시행되었다. 방 법 : 1999년 7월부터 2000년 3월까지 국내 10개 병원에 방문한 정신분열병 및 정신분열형 장애 105명을 대상으로 비대조 개방 임상연구를 시행하였다. 중등도 이상의 정신병 증상을 지니거나, 부작용으로 약물교체가 필요한 경우 또는 기분장애 증상의 치료가 필요한 환자를 대상으로 하였다. 2∼7일간의 약물 배설기간 후에 1일 10mg의 올란자핀을 투여하였고, 이후로 3∼7일 간격으로 용량을 조절하여 총 8주간 투여하였다. 치료 효과는 PANSS(Positive and Negative Syndrome Scale), BPRS(Brief Psychiatric Rating Scale), CGI(Clinical Global Impression-Severity), MADRS(Montgomery-Asberg Depression Rating Scale), HAM-A(Hamilton Rating Scale for Anxiety)로 판정하였다. 안전성 평가는 자발적인 이상반응 보고, 활력징후와 혈액 검사 및 SAS(Simpson-Angus Scale), BARS(Barnes Akathisia Rating Scale), AIMS(Abnormal Involuntary Movement Scale)척도를 이용하였다. 결 과 : 105명중에서 85.7%인 90명이 8주간의 치료를 완료하였다. 평균 최빈 용량은 일일 16.1(±4.7)mg이었고, 종료시점에서의 BPRS전체 점수가 기저 상태에 비해 40%이상 감소된 반응군은 69.5%이었다. PANSS의 양성 증후군과 일반정신병리 뿐만 아니라 음성 증후군 소척도에서도 유의한 감소를 보였으며, 이 감소들은 치료 초기부터 나타나서 8주간 지속되었다. 43.8%의 피험자가 중등도 이상의 우울증상을 같이 지니고 있었으며, 올란자핀에 의해 MADRS와 HAM-A가 유의하게 감소하였다. 치료 기간 동안에 활력징후에는 뚜렷한 변화가 없었으나 체중은 8주간 지속적으로 증가하였다. SAS와 AIMS는 감소하였으며, 정좌불능증을 제외한 나머지 추체외로 증상의 발생비율도 낮았다. 임상적인 증상이나 징후를 동반하지 않는 ALT/SGPT의 상승을 보였지만 프로락틴을 포함하는 대부분의 혈액 검사상 뚜렷한 이상 변화는 없었다. 결 론 ; 비록 본 임상연구가 비대조 개방 임상연구로서 많은 제한점을 가지고 있지만, 올란자핀이 기존 약물에 내약성을 보이거나 기분증상을 동반하는 정신병에 치료 효과가 있었다. 그리고 기존 항정신병약물 치료시에 문제가 되었던 추체외로 증상과 고프르로락틴 혈증을 포함하는 대부분의 부작용면에서 안전성을 보여 주었다. Objective : This multicenter clinical trial was carried out to investigate the efficacy and the safety of olanzpine for the treatment of Korean patients. Mothod : 105 patients with schizophrenia and schizophreniform disorder, visited at 10 mental or university hospitals, had received an open and non-comparative treatment with olanzapine for 8 weeks. Patients had psychotic or depressive symptoms with the severity above moderate degree or intolerable side effects to previous antipsychotics. After a wash-out period of 2-7 days, 10mg olanzapine was prescribed initially to all the patients, and then the dosage could be adjusted within the range of 5-20mg/day of olanzapine by 3-7 days. Results : 90(85.7%) of 105 patients completed the 8-weeks trial and the mean modal dose of olanzapine was 16.1(±4.7%)mg/day. At the end of the trial, 73 patients(69.5%) were classified as responder, which was defined as 40% or more improvement in BPRS(Brief Psychiatric Rating Scale) score comparing to baseline. There was a significant reduction in the scores of PANSS(Positive and Negative Syndrome Scale)and subscales including negative symptom scores and CGI. Also weekly analysis showed that the reductions in scores were kept on for the whole period of the trial. 43.8% of all the patients had depressive symptoms at the baseline and total scores of MADRS(Montgomery-Asberg Depression Rating Scale) and HAM-A(Hamilton Rating Scale for Anxiety) were also reduced after the trials. Vital signs revealed no clinically significant changes but continuous weight gain was observed during the treatment with olanzapine. The scores of SAS(Simpson-Angus Scale) and AIMS(Abnormal Involuntary Movement Scale) for assessing the EPS(extrapyramidal symptoms)and tardive dyskinesia respectively were significantly decreased and only a few patients reported EPS as adverse events. Although mild and clinically non-significant of ALT/SGPT was observed, most laboratory parameters including plasma prolactin level showed to significant changes during the trial. Conclusions : Although this trial had many limitations because it was a non-comparative and open study, olanzapine showed high efficacy on the positive, negative and depressive symptoms in schizophrenia and schizophreniform disorder.In addition to that, olanzapine showed a substantially favorable safety profile, such as low incidence of EPS and hyperprolactinemia.

전경대원에서 발생한 결핵의 임상 양상

문창기,박상준,조민구,김영중,김소연,김윤권,정준오,안석진,김은실,서승오,김지훈,최원제,이윤영,박형기,최규영,김현근 대한감염학회 2006 감염과 화학요법 Vol.38 No.6

Background: The purpose of this study was to document the incidence and clinical characteristics of patients with tuberculosis (TB) in combat and auxiliary police, living in a group, in Korea where the incidence rate of active TB in a general population is higher than in Western countries. Materials and Methods:We retrospectively reviewed the medical records of all tuberculosis patients diagnosed at National Police Hospital from January 2002 through December 2004. Results:In 2002-2004, a total of 156 cases of tuberculosis were identified with the mean (Standard deviation) age of 20.6 (±1.0) years. Of these, 134 (85.9%) patients were registered as new cases, 11 (7.1%) as relapse, 2 (1.3%) as failure, 5 (3.1%) as treatment-after-default cases whereas 4 (2.6 %) patients were not included in any categories. Average annual new TB rate and smear-positive TB rate were 86.5/10^(5) and 17.4/10^(5) person-years, respectively. In 12 of 31 smear-positive cases, time from onset of symptoms to diagnosis was more than 30 days. Two multidrug-resistance TB cases were identified and two suspected outbreak episodes of TB had occurred during 3 years. Conclusion:There was no statistically significant difference in the incidence rate of new cases of TB between the general population aged 20 to 29 years and combat and auxiliary police in Korea. 배경 : 국내 결핵의 발생은 감소 추세에 있으나 여전히 후진국형 발생양상을 보이고 있다. 특히 집단생활을 하는 젊은이들에서 발생하는 결핵의 유행이 간헐적으로 알려지고 있어 우려를 낳고 있다. 이에 저자들은 최근 전경대원의 결핵 발병 양상에 대해 알아보고자 본 연구를 시행하였다. 재료 및 방법 : 2002년 1월부터 2004년 12월까지 3년간 경찰병원에서 결핵 진료를 받은 전경대원의 의무기록을 후향적으로 분석하여 연도별 결핵 신환발생률, 도말양성결핵 신환율, 진단 지연, 집단발병, 다제내성결핵을 조사하였다. 결과 : 연도별 결핵 신환자 수(10만명당 발생추정치)는 2002년 50명(83.6명), 2003년 42명(83.7명), 2004년 42명(93.5명)이었고 도말양성 폐결핵 신환자 수는 2002년 10명(16.7명), 2003년 11명(21.9명), 2004년 6명(13.4명)이었다. 3년간 도말양성 폐결핵 환자는 31명이 있었고 그 중 12명은 호흡기 증상 발생 30일이 지나서 진단을 받았다. 다제내성 결핵은 2명이 있었으며 결핵의 집단발병이 의심되는 사례는 2차례 있었다. 결론 : 전경대원과 20-29세 연령군 일반인의 결핵 신환발생률, 도말양성 폐결핵 발생률의 통계적으로 유의한 차이는 없었다.

분얼형 옥수수의 식물학적 특성과 RAPD 분석에 의한 근연관계

金哲民,李喜鳳,崔現求,鄭載永,金基薰 충남대학교 농업과학연구소 2001 농업과학연구 Vol.28 No.1

분얼형 옥수수 자석계통의 식물학적 특징, 유전적 거리를 조사하여 새로운 교잡종 육성을 위한 기초자료를 얻고자 실험 한 결과를 요약하면 다음과 같다. 1. 개화기에 있어서 조사된 20계통 중 K15가 가장 빨랐고 K07이 가장 늦은 개화소요일수를 보였다. 2. 간장과 착수고는 비슷한 경향으로 보며 K04가 가장 컸고, K15가 가장 작은 것으로 나타났다. 3. 상관 분석 결과 분열수는 개화기 및 출사기와 고도의 정의 상관을 보였으며, 100립중은 개화기, 이삭높이와 고도의 부의 상관을 가지고 있다. 4. RAPD 분석결과 17개의 BAND가 나타났고 이중 1개만이 monomolpic band를 보였으며, primer 당 평균 4개의 loci를 보였다. 5. 유전적 거리와 식물학적 특성간에는 개화기, 분얼수, 엽장과 100립중에서 관련이 있었다. This study was carried out to obtain a basic information for the development of a new corn hybrid with tillers. Materials used in this study were 20 lines having three to four tillers per plant including the PI213749 U S line with non-tillers. These 20 lines were compared for the botanical characteristics and genetic distances were measured using RAPD analysis. Flowering date of the K15 was very earlier. while the K07 was very late in flowering date. Stem height and ear height were similar except for KO4 and the K15. K06, K13 and K19 lines were appeared to be resistant to lodging due to decreased ear height. Number of tillers per plant of lines used were shown three to five on average. KO9 showed the highest kernel yield, while the K08 was low. Among characteristics measured tiller per plant and flowering dates, and silking dates showed a positive correlation, while 100 kernels weight, flowering date and ear height were shown a negative with tillers per plant. A total of 17 bands by RAPD analysis using four per primer were appeared and these lines were classified into three groups, especially the third group could be classified into of four sub-groups.

殺蟲性 結晶蛋白質 特異的인 抗體의 生産

김기윤,이정민,유형진,백길현,권무식 성균관대학교 생명과학자원연구소 1996 生命資源科學硏究 Vol.3 No.1

Bacillus thuringiensis is a gram-positive soil bacterium characterized by its ability to produce crystalline inclusions during sporulation. These parasporal bodies consist of protoxins known as crystal proteins exhibiting highly specific insecticidal activities. The Lepidoptera-specific protoxin, Cry IAc, was purified from E.coli JM103 harboring the cry IAc gene. The gene was isolated from Bacillus thuringiensis kurstaki HD-73 and subcloned into an expression vector, pKK223-3. The recombinant DNA(pOS4201) was transformed into E.coli JM103. The Cry I Ac protoxin overexpressed in the E.coli was isolated from the total proteins by differential solubility. The protoxin were trypsinized to obtain activated toxin. The proteins was resolved on SDS gel. The molecular weights of the pro-and toxin were resistered about 130,000 and 65,000 Daltons respectively. The Cry IAc toxin was used as immunogen to produce anti-Cry IAc antiserum in a rabbit. The immunogen (200㎍/ 200㎕) was mixed with an equal volume of Freund's complete adjuvant, and the mixture was hypodermically injected to the hide back (30 spots) and paws (2 spots) of the rabbit. The immunization was performed four times every two weeks. The dosage of the immunogen was decreased by half, and incomplete adjuvant was substituted for the complete one from the second immunization. The serum was prepared from the blood as described elsewhere. Immunoreactivity of the serum was examined by dot-blotting with the aid of GAR-HRP indirect immunoassay kit. It has been found that the anti-Cry I Ac antiserum recognized nano-gram quantity of antigen under these experimental conditions. The antiserum will he used for the development of immunochemical mean(s) to screen transgenic plants transformed by the insecticidal characteristics of Cry I Ac.