Bacillus sphaericus ts - D1290 치사돌연변이주의 단백질 패턴

이형환,유관희,서정희 한국유전학회 1990 Genes & Genomics Vol.12 No.3

Bacillus sphaericu; ts-D1290 lethal mutant was comparatively characterized with the wild type strain 1593 by the measurements of the biosynthesis of total proteins on the temperature-sensitive cultures at permissive temperature of 30℃ and at nonpermissive-temperature of 42℃. The protein syntheses in the spores of the ts-D1290 and the wild type strain at 30℃ occurred in 6 to 8 hours after inoculation. When the spores of the mutant were cultured at 42℃, they germinated, but the protein synthesis almost did not occur, and the number of viable cells did not increase at 42℃. Vegitative cells after the germination of the wild type spores at 30℃, proteins of 140 and 150 kilodaltons produced within 1 or 2 hours in the cells; however, after a 2 h culture, the amounts of the 150 Kd protein increased. While the 140 kd protein did not produced in the mutant, only the 150 Kd protein produced in the cell, The 155 kd protein produced after a 4 h culture at 30℃.

덕성 함양을 위한 충효행동의 습관화 지도 방안

이형환 慶尙大學校 經營行政大學院 2000 最高管理者課程論文集 Vol.17 No.-

HPLC에 의한 Bacillus thuringiensis 분리균의 내독소결정체 아미노산 조성의 분석

최도숙,오주석,강이석,원남희,차성철,이형환 建國大學校基礎科學硏究所 1999 理學論集 Vol.24 No.-

토양에서 분리한 B. thuringiensis 분리균주들의 내독소결정체 단배질의 조성을 비교 분석하는 것이 목적이었다. B. thuringiensis 분리균주들 (HL-7∼HL-42)의 내독소결정체 층이 형성된 밀도분포는 0.3 g/㎤ ∼0.6 g/㎤이었다. 결정체 단백질을 용해시킨 후에 HPLC분석으로 얻은 아미노산은 14종이었으며, 각 아미노산들의 함유비율은 aspartic acid 6.7∼12.8, glutamic acid 7.0∼12.8, serine 5.9∼12.9, glycine 15.1∼35.5, threonine 3.2∼9.7, arginine 0.5∼3.2, alanine 6.2∼14.6, tyrosine 2.1∼9.0, methionine 0.4∼3.8, valine 3.3∼8.9, phenylalanine 3.3∼8.4, isoleucine 3.1∼7.6, leucine 6.3∼10.9, lysine 2.1∼6.7, mol%의 범위로 분포되었다. glycine이 19.6 mol%로 최고치로 함유되었고, methionine의 함유량은 2.7 mol%로 가장 적었다. nonpolar, polar, acidic 그리고 basic amino acid 순으로 많이 함유되어 있었다. Amino acid compositions of the δ-endotoxin crystal of thirty-six B. thuringiensis isolates (HL-7 to HL-42) were analyzed by high liquid performance chromatography. The crystals of Bacillus thuringiensis isolates were purified by NaBr density-gradient ultracentrifugation. The crystals from the HL-7 to the HL-42 strains contained fourteen different amino acids; aspartic acid, glutamine, serine, glycine, threonine, argnine, isoleucine, leucine, and lysine. The residual values of glycine was highest(19.6%) and that of arginine was lowest(2.7%). The average amounts of the nonpolar amino acid was 42.2%, and that of the nonpolar amino acid was, the acidic amino acid and the basic amino acid.

Escherichia coli RY13의 Auxotrophic Mutant의 분리와 요구성 연구

李炯煥,姜判祚 건국대학교 교육대학원 1982 敎育論叢 Vol.2 No.-

Escherichia coli RY13 were cultivated in Nutrient broth at 37℃ and mutagenized with Nitrosoguanidine. From the mutagenized cells, 47 auxotrophic mutants were selected. Four mutants out of 47 auxotrophs did not require any amino acids at all, but 43 mutants out of them required all kinds of amino acids.

미생물 살충제의 특성과 전망(1) -곤충바이러스의 연구현황과 전망-

이형환 한국작물보호협회 1984 자연과 농업 Vol.5 No.11

소(牛)성장호르몬 유전자 클로닝의 기초 연구

李炯煥 建國大學校基礎科學硏究所 1987 理學論集 Vol.12 No.-

소(牛)의 뇌하수체전엽에서 분비되는 소의 성장호르몬을 얻기 위하여 전엽 추출물을 PAGE 후에 젤의 각 밴드를 추출한 다음 RIA 방법을 이용하여 ??항체와 반응시켜 소의 성장호르몬양을 측정하였다. 측정결과 뇌하수체 gram당 6.69㎍의 성장호르몬이 있는 것으로 나타났다. 뇌하수체 분쇄물에서 RNA를 분리하기 위하여 agarose gel 전기영동을 한 결과 분자량이 0.08Kb에서 21.22Kbs의 크기를 갖는 8개의 밴드가 나타났고, 분쇄물을 다시 oligo dT 컬럼을 통과하여 총 mRNA를 얻었다. 이 컬럼을 0.1M NaCl과 NaCl이 없는 완충액으로 유출시킨 결과는 NaCl이 없는 유출액 중에 성장호르몬 mRNA의 양이 0.1M NaCl 유출액에서 보다 7배 정도가 많았다. mRNA 분획을 AMV reverse transcriptase을 이용하여 cDNA을 합성한 결과는 100㎕의 합성반응액에서 약 0.2㎍의 DNA을 얻었다. Bovine growth hormones were extracted from bovine pituitary glands and then measured by PAGE and RIA. Bovine growth hormone was reacted with ?? antibody. There were 6.69 ㎍ per gram of bovine pituitary gland. Growth hormone RNAs were extracted from the gland, which were electrophorized on 1.0 % agarose gel. There were formed about 8 different bands ranging from 0.08Kb to 21.22 Kbs. Total mRNAs were obtained by oligo dT chromatography, which were in vitro translated. In the eluent by elution buffer not contained NaCl, growth hormone mRNAs were presented 7 times more than 0.1 NaCl eluent. The mRNA eluent was used for template for cDNA synthesis by AMV reverse transcriptase. From 100㎕ of the reactant,about 0.2㎍ of cDNA were obtained.

Escherichia coli의 Temperature-sensitive Mutants의 誘導分離

李炯煥 建國大學校基礎科學硏究所 1983 理學論集 Vol.8 No.-

Escherichia coli RY13 K-153 균주를 37℃의 영양배지에서 배양하면서 Nitrosoguanidine을 처리하여 돌연변이를 유도한 후에 한천영양평판배지에 콜로니를 형성시키었다. 이들 콜로니들이 42℃에서 온도민감성을 나타내는지를 검사하여 온도민감성 돌연변이 균주를 선정하였다. 총 21개의 온도민감성 돌연변이 균주를 선택하여 42℃의 제한온도에서 성장에 대하여 조사를 한 결과는 다음과 같다. 21개의 돌연변이 균주 중에서 10개는 42℃에서 전혀 성장을 안하였으며, 11개는 약간씩 성장을 한 것으로 나타났다. 이러한 성격으로 보아서 전자는 치사군, 그리고 후자는 성장저조군으로 나누었다. 정상형과 돌연변이 균주에 대한 항생제 저항성을 조사한 바, 정상형은 ampicillin, chloramphenicol, erythromycin, penicillin, streptomycin과 tetracycline에 저항성을 나타냈고, cephalothin과 kanamycin에는 민감했다. ts-N56, -N104와 -N25는 kanamycin에 민감성을 나타냈고, ts-N63과 ts-N87는 ampicillin에 저항성을 나타냈으며, ts-N132, -N25와 -N130은 tetracycline에 저항성을, ts-N25와 -N130은 erythromycin 민감성으로 변했다. Escherichia coli RY13 K-153 was cultivated in nutrient broth at 37℃ and then mutagenized with N'-methyl-N'-nitro-N-nitrosoguanidine at 37℃. Temperature-sensitivity was checked at 37℃ (permissive temperature) and at 42℃ (nonpermissive temperature). Twenty-one temperature-sensitive mutants were isolated from the mutagenized cells and characterized at 42℃. The mutants were classified into two groups: lethal group and non-lethal group. Ten mutants out of twenty-one belong to the lethal group, and eleven mutants out of the twenty-one mutants belong to the non-lethal group. The wild type and the temperature-sensitive mutants were examined for antibiotic susceptibility at 37℃. The wild type was resistant to ampicillin, chloramphenicol, erythromycin, penicillin, streptomycin and tetracycline, and was sensitive to cephalothin and kanamycin. The ts-N56, -N104 and -N25 were changed from sensitive phase to resistant phase in kanamycin, the ts-N63 and ts-N87 were changed to ampicillin resistance. The ts-N132, -N25 and -N130 were shifted to tetracycline resistance. The ts-N25 and -N130 were changed to erythromycin sensitive mutant.

녹용 추출액이 대장균 항체생산에 미치는 영향

이형환,제갈승주 한국미생물학회 1983 미생물학회지 Vol.21 No.1

Commercial deerhorns were extracted in boiling water, the deerhorn extracts were per os introduced into rabbits, and then the effects of the extracts on the antibody productions aginst Escherichia coli antigens were investigated for 4 weeks. The experimental rabbits were divided into 4 groups ; control, only deerhorn, only antigen, and antigen plus deerhorn treated groups. The effects of the treatments were measured by counting the number of blood cells, weighing, and immunoelectrophoresis. The rabbits' body weights gained up to 185% in the deerhorn group, and the other groups gained nearly 120%. The numbers of red blood cells in the antigen plus deerhorn group increased somewhat. However, the numbers of leucocytes gradually increased after one week in the antigen group, and at 4 weeks increased up to 290%. In the antigen plus deerhorn group the numbers of leucocytes increased suddenly up to 189% at one week, but after one week the numbers recovered to normal state. Strangely in the deerhorn group the numbers decreased up to 40%. The amounts of serum globulin increased in the control after one week, but maintained about 130%. In the deerhorn group the amounts increased like the control, but after 4 weeks increased up to 175%. In the antigen group the amounts were not changed until 2 weeks, but after 3 weeks abruptly increased over 175%. In the deerhorn plus antigen group the amounts increased gradually up to 262% until 3 weeks, after 3 and 4 weeks the amounts did not increase. The amounts of serum .gamma.-globulin decreased in the control group, and in the deerhorn group and antigen group the amounts did not change until 2 weeks, but after 3 weeks abruptly increased over 175%. In the deerhorn plus antigen group the amounts increased gradually up to 262% until 3 weeks, after 3 and 4 weeks the amounts did not increase. The amounts of serum ${\gamma}-globulin$ decreased in the control group, and in the deerhorn group and antigen group the amounts did not change until 3 weeks, but after 4 week the amounts slightly increased up to 110%. However, the amounts in the deerhorn plus antigen group did not change until 2 weeks, but after 2 weeks abruptly increased up to 174%. The recognized immunoglobulins were IgG and IgM, and the enhanced immunoglobulin was IgG.

슬개골 골절의 수술적 치료

이형환,손성근,이규열,김성수 대한골절학회 1996 대한골절학회지 Vol.9 No.3

The patella, the largest human sesamoid bone, is important functional component of the knee extensor mechanism and its treatment is considered to be very important because the fracture involvement to the articular surface occures in most of the cases. We report 40 cases of fractures of patella treated with operative methods from January 1990 to June 1995 at the department of orthopaedic surgery, Dong-A university hospital. The results are as follows. 1. Among 40 cases, 29 cases were male and 24 cases were belong to 3rd decade and 4th decade. 2. The most common cause of injury was slip down or fall down. 3. The pattenrs of fracture were transeverse in 2 l cases, vertical in 2 and comminuted in 17. 4. The methods of surgical treatment were modified tension band wiring in 33 cases, circumferential wiring in 2, partial patellectomy in 4 and total patellectomy in l. 5. In patient treated with modified tension band wiring, range of motion of the knee was l28and mean duration of bone union was 9.2 weeks. 6. Complications were limitation of range of motion in 3 cases, refracture by slip down in 1 case and wire migration in 1 case.

Bacillus thuringiensis var. darmstadiensis 와 var. sotto의 내독소와 Bacillus thuringiensis var. kurstaki가 생산하는 아밀라제의 연구

이형환,김삼찬,황광현,임창로,조용칠,정인명 건국대학교 교육대학원 1991 敎育論叢 Vol.15 No.-

Endotoxin crystals produced by B.thuringiensis var. darmstadiensis cultured in modified GYS medium over 72 h were ovoidal in shape and separated using Renografin gradient centrifugation. The crystal protein bands were observed. When B.thuringiensis var sotto was grown in the modified GYS broth, it entered stationary phase at 8-9 h after inoculation. Proteinaceous crystals were banded at the position in 40% of Renografin gradient. The solubilized crystal proteins were formed by two bands, 130 Kd and 68 Kd in the SDS-PAGE. One plasmid was observed in B.thuringiensis var darmstadiensis and no plasmid in var. sotto in this study. Enzyme activity of amylase produced B.thuringiensis var kurstaki strain in the basal medium was 0.4 units per ml, and was enhanced to 0.6 units per ml in the basal medium contained soluble starch, Ca^++, Mg^++ and Mn^++ ions. Amylase production medium containing soluble starch was suitable for the amylase production and the highest activity appeared at 4-6 h after cultivation at 32℃. The amylase activity obtained by ethanol precipitation was 0.20 units per ml in 0.1 M phosphate buffer. The band of the molecular weight of 50 Kd was observed by SDS-PAGE analysis. Km value of the amylase for the soluble starch was 6.80 mg per ml.