장독성 대장균 eKT - 53균주의 내열성 장독소의 성질

도대홍(Dae-Hong Do),김교창(Kyo-Chang Kim),김도영(Do-Young Kim) 한국식품영양과학회 1991 한국식품영양과학회지 Vol.20 No.6

형질전환균주 eKT-53(ST^+ LT^-)를 사용하여 succinate salts 배지에 배양한 배양상층액으로 부터 분리 정제한 내열성장독소(ST ; heat-stable enterotoxin)의 몇가지 성질에 대하여 조사하였다. 분리 정제된 ST의 내열성은 100℃에서 30분 이상 열처리하였을 때 ST 활성이 소실되었다. pH에 대한 안정성은 pH 2~10에서는 대단히 안정하였지만 pH 1과 pH 12에서는 불안정하였다. 또한 α-amylase(Bacillus subtilis) 와 pepsin(porcine)에 대해서 매우 안정하여 활성변화가 없었으나 disulfide 환원제를 처리할 경우 쉽게 활성이 소실되었다. 이러한 ST의 분자량은 약 4,200이었고 pI값은 4.0이였다. Heat-stable enterotoxin(ST) from enterotoxigenic E. coli eKT-53(ST+ LT-, transformant from isolate KM-7) that was produced in succinate salts medium. The culture supernatant(crude ST) was purifed by multipled steps and investigated some characterization of the ST. The heat-stability of purified ST activity was completely lost by treating at 100℃ for 30minutes. ST activity was lost by treatment at pH 1 and 12 conditions, while the activity was not reduced by treatment at pH 2~10, and then the α-amylase and pepsin was not decreased activity but disulfide reducing agents was lost the activity. The molecular weight of the purified ST was approximately 4, 200, the isoelectric point was about 4.0.

식품폐수의 영양원 분해를 위한 Pseudomonas sp.의 분리 및 특성에 관한 연구

한재석(HAN Jae-Seok),도대홍(DO Dae-Hong) 한국환경관리학회 1997 環境管理學會誌 Vol.4 No.-

Aspergillus fumigatus가 생산하는 cellulase에 관한 연구

도대홍 충북대학 대학원 1981 大學院論文集 Vol.7 No.-

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쥐의 心臟筋肉 細胞로부터 mRNA 分離 精製및 cDNA 合成

都大洪,金敎昌 충북대학교 농업과학기술연구소 1987 農業科學硏究 Vol.5 No.2

mRNA from Atrial muscle of rat heart has been purified poly(A)+RNA, and its physical properties have been examined. Isolation was achieved by the use of guanidinium extraction method to isolate cellular RNA. Purification was achieved by the use of oligo(dT)-cellulose chromatography to separate poly(A) +RNA from ribosomal RNA, Poly(A) +RNA was translation in a cell-free reticulocyte system. Purified poly(A) +RNA was synthesized a double strand DNA by using reverse transciptase, polymerase arid T4 ligase. The extracted mRNA was obtained approximately 5mg against 20g sample tissue (rat heart). The purification fold was 40-fold, and final purified poly(A) +RNA in orig(dT)-cellulose chromatography was taken 2.4% of the initial mRNA. When 1.0㎍ of poly(A) +RNA was added to a cell-free system in 25㎕ reaction volumn, the complex protein synthesizing ability was maximized. After purifying the poly(A):RNA it was synthesized to a single and then a double strand DNA. Its conversion rate was figured 0.38% for a single strand, and 0.356% for a double strand DNA.

쥐의 심장근육 세포로부터 mRNA 분리 정렬 및 cDNA 합성

도대홍, 김교창 忠北大學校 遺傳工學硏究所 1988 遺傳工學硏究 Vol.2 No.1

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돼지에서 由來한 耐熱性 腸毒素生産 大陽菌의 分離 및 判定

都大洪,金敎昌,金道榮 충북대학교 농업과학기술연구소 1987 農業科學硏究 Vol.5 No.2

Enterotoxigenic E.coli is the one of major causative occured to the infantile swine diarrhea. A total of 189 isolated idem assumed E. coli, Salmonella and Shigella was isolated from swine in Kyong-Gi and Chung-Cheong province. When enterotoxigenic E. coli was infected to infant swine, these adhered to the mucosa of small intestine with pili and excrete enterotoxin, which caused the diarrhea. The detection of a heat-stable enterotoxin(ST) produced of isolated E.coli was porformed by infant mouse assay, and was identified pili by the aggutination test of rbc suspention. The plasmid DNA profile was investigated by the agarose gel electrophoresis. The results of assay to enterotoxigenic E.cofi summarized as follow. Isolation ratio of enterotoxigenic E.coli among total of 45 isolated strains was taken 56.52% during summer. Four strains(KS-4, KM-7, KM-12, CM-7) of high produced ST were showed the G/B ratio above 1.10 by infant mouse assay. Four strains of high produced enterotoxigenic E.coli was isolated from swine with diarrhea, and 3 strains (KS-4, KM-7, KM-12) were proved to possess K88 and K99 antigens, or K88 antigen. The patters of plasmid DNA was contained 4.5kb plasmids encoded for ST. The antibiotics sensitivity patters of cultured enterotoxigenic E.coli were showed a tendency to be resistant against tetracycline and streptomycin.

病原性 大腸菌이 생성하는 耐熱性 腸毒素의 抗體 생성조건 및 中和效果

金敎昌,都大洪,鄭埈永 충북대학교 농업과학기술연구소 1993 農業科學硏究 Vol.11 No.1

The heat-stable enterotoxin(ST) producing Escherichia coli strains were isolated from the swine. The purified ST was immunized with ST-BSA conjugate and obtained antiserum from a rabbit (New Zealand Whit). A pathogenic toidity was neutralized with anti-rabbit serum. When antiserum obtained from rabbits immunized with ST- BSA antigens. It was production the most of antiserum at the fourth immunization (21 days). Obatined antisera neutralized the toxic activity of purified 4 mouse units to 1 : 32 of antisera dilution folds.

채소연부병균 Erwinia herbicola의 생육억제균 분리 및 특성

김교창,도대홍,김도영 한국식품영양학회 1996 韓國食品營養學會誌 Vol.9 No.3

Erwinia herbicola(EH) 채소 연부병 방제를 위하여 충북 청원군, 진천군 및 충남 연기군 일원의 채소 경작지 토양으로부터 형광성을 나타내는 Pseudomonas속 1,196균주를 선발하고, 연부병균 Erwinia herbicola에 대한 생육억제력과 생물학적 검정으로 발병억제력이 강한 2균주를 선발하여 Pseudomonas fluorescens S4(PS4)와 Pseudomonas fluorescens S65 (PS65)로 동정하였다. 배춧잎을 이용하여 분리길항균의 연부병균에 대한 생물학적 억제력을 확인한 결과 상당한 발병억제력을 확인하였다. 이들 두 균주가 가장 높은 길항력을 나타내는 조건은 배양 초기배지의 pH를 7.0으로하여 25℃에서 3일간 배양했을 때였다. 화학농약 및 항생물질에 대해 2균주 모두 benomyl, propamocab hydrochloride, fosethyl-Al-folpet, vancomycin, penicillin 및 lincomycin에 내성을 나타냈고, PS4균주는 erythromycin에 대해서도 내성을 나타냈다. For the selection of powerful antagonistic bacterium for biological control of Erwinia sp. causing vegetable soft rot, two excellent strains(S4, S65) were selected from 1,196 strains of bacteria which were isolated from rhizospere in vegetable root rot suppressive soil. Selected 2 strains were identified to be a species to Pseudomonas fluorescens S4(PS4) and Pseudomonas fluorescens S65 (PS65). The highest of inhibitory activity was produced in 523 synthetic broth medium at pH 7.0 and 25℃ during 3 day culture. The isolate strains were resistant to the agricultural chemicals such as benomyl, proamocab hydrochloride and fosethyl-Al-folpet, and the antibiotics such as vancomycin, penicillin and lincomycin, only PS4 was resistant to erythromycin.

땅두릅뿌리를 이용한 혼합음료가 당뇨 및 고혈압 환자의 혈압과 혈액 성상에 미치는 영향

최미숙,도대홍,최도점 한국식품영양학회 2002 韓國食品營養學會誌 Vol.15 No.2

충청북도 청주시 흥덕구에 거주하고 있는 24명의 독거 노인 (남자 10명, 여자 14명)으로 당뇨병 환자이고 경계성 고혈압 이상에 해당되며, 60세 이상인 노인을 대상으로 혈압과 혈청학적인 검사를 실시하여 얻은 결과는 다음과 같다. 1. 수축기 혈압은 3주에 1주보다 A군에서는 4.9% 상승하였고 B군은 0.3%가 증가되었으며 C군에서는 5.9%가 감소되었다. 각 군의 3주 평균은 A군 158.3±24.3, B군 149.1±19.2, C군 143.9±5.1로 C군, B군, A군 순으로 통계적으로 유의하게 낮은 것으로 나타났다. 이완기 혈압은 1주에 비해 3주에 A군에서는 3.7% 증가하였고 B군은 0.3% 감소하였으며 C군에서는 1.5% 증가하였다. 각 군의 3주 평균은 A군 93.8±11.3, B군 93.2±11.2, C군 86.9±6.2로 C군, B군, A군 순으로 낮게 나타났다. 이 결과에 의하면 두릅음료가 수축기와 이완기 혈압 조절에 기여하는 것으로 보인다. 2. 혈당은 3주에 1주보다 A군에서는 0.5%가 증가하였고 B와 C군에서는 각각 13.9%와 17.5%가 감소되어 두릅음료가 혈당저하에 효과적인 것으로 나타났다. 3. 중성지방은 1주에 비해 3주에 A군에서 11.6% 증가되었고 B군에서는 5.0% 감소하였으며 C군에서는 3.8%가 증가되었다. 총콜레스테롤은 1주보다 3주에 A군에서는 2.3% 증가되었고 B와 C군에서는 각각 8.7%와 14.5%가 감소하였다. HDL-콜레스테롤은 A와 B군에서는 각각 4.8%와 13.2%가 낮아졌고 C군에서는 19.6%가 증가되었으며 LDL-콜레스테롤은 1주보다 3주에 A군에서 1.9% 상승하였으나 B와 C군에서는 각각 8.3%와 21.1%가 저하되었다. 이 실험 결과에 의하면 두릅음료가 총 콜레스테롤과 LDL-콜레스테롤 저하와 HDL-콜레스테롤 증가에 효과적인 것으로 나타났고 중성지방의 조절에는 효과적이지 못한 것으로 나타났다. 4. GOT는 3주에 1주에 비해서 A군은 16.4% 감소하였고 B군은 14.9% 증가하였으며 C군은 14.7%가 저하되었다. GPT는 3주에 1주보다 A군 1.0%, B군 43.0%, C군은 43.0%의 증가를 나타내었다. 총 빌리루빈은 3주에 1주에 비해 A군에서는 변화가 거의 없었고, B군에선 25% 감소를 보였으며 C군은 50%가 증가되어 GOT, GPT 및 총 빌리루빈의 농도는 두릅음료에 의해 저하되지 않는 것으로 나타났다. 5. 총 단백질은 3주째 1주보다 A군은 13.6%, B군은 8.6%, C군은 11.8%가 증가되었고 알부민은 3주째 1주에 비해 A와 C군은 증가가 없었고, B군에서만 11.6% 감소하였으며 혈중요소질소는 1주보다 3주에 A와 B군은 각각 6.3%와 14%가 감소되었고 C군에서는 2.2%가 증가되었으며 요산은 3주에 1주보다 A와 C군에서 각각 20.4%와 11.1%가 감소하였고 B군에서만 14.3%가 증가하여 두릅음료의 효과가 총 단백질, 알부민, 혈중요소질소 및 요산에서는 뚜렷하게 나타나지 않았다. Studies were carried out to observe the influence of the Aralia continentalis Kitagawa root on blood pressure and blood constituents of the 24 hypertensive elderly (10 men and 14 women aged over 60). Total subjects were divided into three groups according to taking antihypertensive medications (A), antihypertensive medications and mixing beverages with the Aralia continentalis Kitagawa root (b) or mixing beverages with Aralia continentalis Kitagawa root (C). The systolic blood pressure of C group was significantly lower than that of A and B groups. The diastolic blood pressure of C group was lower than A and B groups. The blood glucose level of C group was a little lower than that of A and B groups. The level of TG increased more in A group than C group and decreased in B group. The levels of T-cholesterol and LDL-cholesterol lowered in B and C groups and HDL-cholesterol level was higher in C group. The GOT, GPT and T-bilirubin levels of C group were significantly higher than others. Mixing beverages with Aralia continentalis Kitagawa root did not affect on blood protein and non-protein nitrogens. In conclusion, mixing beverages with Aralia continentalis Kitagawa root had an effect on lowering blood pressure, blood glucose, T-cholesterol and LDL-cholesterol concentrations.

돼지에서 유래한 병원성 대장균의 내열성 장독소 생산유전자의 Cloning 및 발현

김교창,도대홍 한국식품위생안전성학회 1991 한국식품위생안전성학회지 Vol.6 No.3

Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin(ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be s maker for identification of the enterotoxingeic E. coli from non pathogenic E. coli. The isolate of enterotoxigenic E. coli was isolated from swine during 1989 yesr(from 5 to 10 month) in the Kyong-gi and Chung-Cbeong provinces, and three strsins(KM-4, KM-7 and KM-12) was selected from 189 isolates of ST producing E. coli. The detection of a ST produced of the isolated E. coli was performed by the infant mouse assay(IMA). This study was designed to know optimal conditions for the production of the ST and the molecular properties of plasmids of the enterotoxigenic E. coli. Amount of ST produced were the most at initial pH 8.5-r9.0 of succinate salts medium culture. The cultural time of the same medium was accumulated the highest level of ST was at the 14 to 16 hours, and then stationary phase wss at the 20 hours. From this experiment the KM-7 strain was selected among ST producing strains by IMA. Partial plasmid-curing experiment was done to select plasmid encoding for ST among other plasmids and then comparing the plasmid pattern of ST producing slrain(KM-7) with those of other ST non-producing strains, it is found that ST gene exists on the about 80 Kbp plssmid. Each fragment of this plasmid digested with EcoRi was ligated to vector pBR 322 and transformed into E. coli K-12. A clone producing ST(eKT 53) was selected by IMA. The EcoRI digestion pattern of the isolated plasmid(pKD 37) from the ST producing clone it is indicated that the size of the inserted fragment in eKT 53 strain is 16 Kbp. The cultured supernatant of eKT 53 strain was positive result of ST production in IMA.