• 식물병연구
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• 제9권2호
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• pp.89-93
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• 2003

남해지역의 원원종 종서 생산 포장에서 '추백' 품종에 나타난 엽맥투명 및 매우 약한 모자이크 증상을 나타내는 감자 잎에서 담배 모자이크 바이러스(TMV)를 분리하였다. 이 바이러스((TMV-St))는 생물학적, 혈청학적 유연관계 및 외피단백질의 염기서열 등을 통해 기존에 보고된 다른 tobamovires와 비교하였다. TMV-St는 5개의 지표식물 반응에서 토마토, 고추, 가지 등과 같은 가지과 작물에 경제적 피해를 주고 있는 TMV-U1, Pepper mild mottle virus(PMMoV) 및 Tomato mosaic virus(ToMV)와는 다른 기주 반응을 보였다. 특히 즙액접종에 의한 기주의 반응은 C.murale 접종엽과 상엽 모두에서 퇴록반점을 보였으며, C. murale, G. globosa, N.rustica 그리고 N. tabacum ce. Samsun nn 등 4가지 지표식물로 이들 바이러스 계통을 구분할 수 있었다. 혈청학적 검정에서 TMV-St는 TMV-U1, PMMoV 그리고 ToMV와의 반응에서 도두 뚜렷한 침강선을 형성하였다. TMV-St의 외피단백질은 477개의 염기서열로 되어 있으며, 이는 TMV-U1의 염기서열과 매우 유사하였다.

• 식물병연구
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• 제9권3호
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• pp.107-115
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• 2003

경상북도 안동, 의성, 군위, 대구의 장마 재배포장에서 모자이크와 괴저반점을 보이는 장마잎을 채집하였다. 이들 이병 잎 조직을 시료로 하여 D N 법 및 ISEM법에 의해 투과전자현미경으로 바이러스 입자를 관찰한 결과, 각각의 시료에서 ChYNMV 항혈청에 반응한 660nm의 사상형 입자가 확인되었다. 장마 잎에서 부분 정제한 바이러스에서 RNA를 추출하여 이것을 주형으로 ChYNMV 특이적 프라이머와 oligo-dT 프라이머를 이용하여 외피단백질 유전자와 3‘-말단 비전사부위를 포함하는 약 1.2kbp의 3’-말단을 증폭하였다. 외피단백질 아미노산서열은 Macluravirus로 알려진 ChYNMV (A B044386)와 97.9%의 상동성을 보여 장마에서 분리한 바이러스를 ChYNMV로 동정하였다. ChYNMV와 다른 Macluavirus의 외피 단백질 아미노산서열을 비교한 결과, M말단 영역에서 가장 많은 변이를 확인하였고 Macluravirus 속에는 잘 보존된 영역이 존재하였다.

• 대한물리의학회지
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• 제9권1호
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• pp.45-53
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• 2014

PURPOSE: In this study, we tried to find out what kind of exercise was more effective in sleep disorder by comparing melatonin in blood after applying low intensity with high intensity exercise to sleep disordered rats induced by experiment. METHODS: We used male Sprague-Dawley rats which were 8weeks old and weighted 300g. They were supplied with water and food without any restriction. We kept the room temperature at $25^{\circ}C$ and controld the length of day and night in 12 hours blocks, respectively. We divided the rats 60 into 2 groups. To one group we applied low intensity exercise, and to the other we applied high intensity exercise for 15minutes per day over a period of 4 weeks. We extracted the blood from abdominal aorta before, after exercise, moved into EDTA tube, performed centrifugation. We decanted the serum $200{\mu}l$ from the blood into microcentrifuge tube by samples and moved into polypropylene culture tubes with micro pipette. We split enzyme solution $50{\mu}l$ into the tubes with melatonin direct kits and make them react at $37^{\circ}C$ for 2 hours. We split assay buffer $50{\mu}l$ into each tube and mixed melatonin tracer $50{\mu}l$ and melatonin antiserum $50{\mu}l$, respectively. After we made them react in room temperature, we decanted the superficial layer with a centrifuge and measured the activity for 1 minute by competitive method with ${\gamma}$-counter equipment. We draw a standard curve through logit-log graph with CPM(counts per minute) and counted the melatonin by B/B0. We conducted independent t-test to examine the homogeneous of melatonin value of before low-intensity and high-intensity exercise. We performed paired t-test to compare before and after low-intensity and high-intensity exercise, respectively. We carried out independent t-test to compare melatonin value after low-intensity and high-intensity exercise. Significance level was .05. RESULTS: The results were as follows; firstly melatonin was more increased in the group who was exposed to high intensity exercise when we compared before to after high and low intensity exercise, respectively. Secondly, high intensity exercise was more effective than low intensity exercise when we compared the two. CONCLUSION: In conclusion, secretion of melatonin which is the material of sleep improvement could be promoted by high intensity exercise. Low intensity exercise acted as a stress rather than improving sleep and had a negative effect on the secretion of melatonin because the melatonin was affected by stress.

• Journal of Nutrition and Health
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• 제32권6호
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• pp.628-636
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• 1999

There is a marked increase in geriatric disease, especially liver disease, due to the continuous increase in alcohol and fat consumption. Since the fatty liver, induced by alcohol or fat, is basically from abnormalities in the lipid metabolism, it is possible that fatty acid binding protein(FABP) which is related to the fatty acid metabolism may also be abnormal in these livers. FABP is a small molecular weight protein family present in cytosol in high concentration. It has been proposed as a fatty acid transfer protein and as a binding protein responsible for controlling intracellular free fatty acid concentration. In this research, we have examined the relationship between liver FABP and fatty liver induced by alcohol or high cholesterol diet. Rats were fed one of either semipurified liquid diets; control diet containing 65% carbohydrate, 20% protein, and 15% fat or high cholesterol diet containing 1%(w/w) cholesterol or alcohol diet containing 37% of alcohol instead of carbohydrate. After 5 weeks of feeding period, all rats received commercial chow diet for 5 weeks to examine recovery effect. Liver and blood samples were collected at 0, 1, 3, 5 and 10 weeks to analyze lipid compositions. FABP was purified from liver cytosol and injected to rabbit to obtain antiserum. Liver FABP amount was determined by SDS-PAGE and western blotting methods. Fatty acid binding capacity was determined by binding of 14Cpalmitate with the delipidated liver cytosol. Consumption of alcohol increased serum cholesterol, triglyceride concentration and decreased HDL-cholesterol concentration after 5 weeks. Serum apolipoprotein B concentration increased after 3 weeks and LDL-cholesterol and apolipoprotein A concentration changed after 1 week. Liver cholesterol and triglyceride concentration increased after 3 weeks. Consumption of high cholesterol diet changed liver and serum lipid composition after 3 weeks. Swiching to normal diet for 5 weeks did not normalize most of lipid composition in serum and liver except serum and liver except serum cholesterol, triglyceride and liver cholesterol. Liver cytosol FABP content and the fatty acid binding capacity decreased dramatically after 1 week with alcohol consumption. This results indicate that FABP content changes before the changes before the changes of blood or liver lipid composition, suggesting changes of FABP may cause development of the fatty liver induced by alcohol and can be used as an index of detecting a early development of fatty liver.

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.912-918
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• 2003

Mitotic centromere-associated kinesin (MCAK), which is a novel kinesin with a central motor domain, is believed to playa role in mitotic segregation of chromosome during the M phase of the cell cycle. In the present study, it is shown that a rabbit polyclonal antibody has been produced using the N-terminal region (187 aa) of human MCAK expressed in E. coli as the antigen. To express the N-terminal region in E. coli, the MCAK cDNA fragment encoding N-terminal 187 aa was obtained by PCR and was then inserted into the pET 3d expression vector. Molecular mass of the N-terminal region overexpressed in the presence of IPTG was 23.2 kDa on SDS-PAGE, and the protein was insoluble and mainly localized in the inclusion body that could be easily purified from the other cellular proteins. The N-terminal region was purified by electro-elution from the gel after the inclusion body was resolved on the SDS-PAGE. The antiserum obtained after tertiary immunization with the purified protein specifically recognized HsMCAK when subjected to Western blot analysis, and showed a fluctuation of the protein level during the cell cycle of human Jurkat T cells. Synchronization of the cell-cycle progression required for recovery of cells at a specific stage of the cell cycle was performed by either hydroxyurea or nocadazole, and subsequent release from each blocking at 2, 4, and 7 h. Northern and Western analyses revealed that both mRNA and protein of HsMCAK reached a maximum level in the S phase and declined to a basal level in the G1 phase. These results indicate that a polyclonal antibody raised against the N-terminal region (187 aa) of HsMCAK, overexpressed in E. coli, specifically detects HsMCAK (81 kDa), and it can analyze the differential expression of HsMCAK protein during the cell cycle.

• 한국가축번식학회지
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• 제13권2호
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• pp.113-120
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• 1989

In order to determine the sex of preimplantation embryos prior to transfer in cattle, a series of experiments were carried out using 45 Holstein donor cows to examine the ovarian response on the gonadotropin and PGF2${\alpha}$, and the morphology of fresh embryos or frozen/thawed embryos after deep freezing at -196$^{\circ}C$. The sexing of embryos treated with the medium containing H-Y antiserum(10%, v/v) and FITC anti-mouse IgG(10%, v/v) were analysed by chromosomal analysis, and the sex of the embryos which survived were ascertain after delivering the pups. The results obtained were summarized as follows ; 1. The average number of developed follicle and corpus luteum per cow were 13.5 and 8.1, and the ovalation rate was 60.1%. 2. Of 220-ova recovered, 75(34.1%) were morula and 91(41.4%) were blastocyst, and the morphological normal and abnormal rate of ova recovered were 75.5% and 24.5%, respectively. 3. Of 39 frozen/thawed embryos, the scores of normal morula and blastocyst, after thawing were 79.2%(19/24) and 73.3%(11/15). The average rate of frozen/thawed embryos which appeared morphologically normal post thawing was 76.9%(30/39). 4. The sex ratio was measured using the embryos treated with immunofluorescence assay to examine the relationship between embryo developmental stage, sex ratio of morula stage embryo was 42.2%(19/45) fluorescing and 57.8%(26/45) non-fluorescing, on the other hand, the ratio switched to 46.8%(29/62) fluorescing and 53.2%(33/62) non-fluorescing embryo in blastocyst stage. The sex ratio was also measured between fresh and frozen/thawed embryos, fresh and frozen/thawed treated embryos were indicated 45.8%(38/83) fluorescing, 54.2%(45/83) non-fluorescing and 41.7%(10/24) fluorescing, 58.3%(14/24) non-fluorescing. This trend indicated the approximal sex ratio was 1 : 1. 5. The result of karyotype test showed the successful rate of sexing embryo is fluorescing and non-fluorescing was 21.2%(7/33) and 29.6%(8/27). The female to male ratio within 33 fluorescing was 28.6 : 71.4, and the ratio of 27 non-fluorescing embryos was 87.7 : 12.5. 6. Of the embryo transferred after assignment of H-Y phenotype, five of the fluorescing embryos survived to term, all was males. Whereas six non-fluorescing embryos also survived to term and the sexes of the calves were 1 male 5 female.

• Journal of Microbiology and Biotechnology
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• 제25권6호
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• pp.771-781
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• 2015

To examine if the molecular chaperone DnaK operon proteins of Streptococcus suis type 2 (SS2) are involved in adhesion to host cells, the abundance values of these proteins from the surface of two SS2 strains of different adhesion capability were compared. Their roles in growth and adhesion to human laryngeal epithelial cell line HEp-2 cells were investigated on SS2 strain HA9801 and its mutants with DnaK operon genes partially knocked-out (PKO mutant) under heat stress. The major difference was that DnaJ was more abundant in strain HA9801 than in strain JX0811. Pretreatment of the bacteria with hyperimmune sera to DnaJ, but not with those to other proteins, could significantly reduce SS2 adhesion to HEp-2 cells. PKO of dnaJ g ene resulted in decreased SS2 growth at 37℃ and 42℃, and reduced its adhesion to HEp-2 cells. The wild-type strain stressed at 42℃ had increased expression of DnaJ on its surface and elevated adhesion to HEp-2 cells, which was also inhibitable by DnaJ specific antiserum. These results indicate that the DnaJ of S. suis type 2 is important not only for thermotolerance but also for adhesion to host cells. Because DnaJ expression is increased upon temperature upshift with increased exposure on the bacterial surface, the febrile conditions of the cases with systemic infections might help facilitate bacterial adhesion to host cells. DnaJ could be one of the potential candidates as a subunit vaccine because of its good immunogenicity.

• 한국잠사곤충학회지
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• 제31권2호
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• pp.82-90
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• 1989

분리.정제한 전잠 vitellin의 분자량추정, 당 및 지질성분의 존재유무, 전기현미경적인 형태 및 크기, 아미노산 조성, 내열성 및 여러 가지 특성을 조사하였다. 1. 분리.정제된 천잠의 vitellin은 전기영동상의 이동도가 다른 large 및 small subunit로 되어 있다. 2. 천잠 vitellin의 분자량은 약 450,000d로 추정되었고, large subunit와 small subunit는 각각 174,000d와 44,000d로 추정되었다. 3. Coomassie brilliant blue, sudan black B 및 PAS 염색에 의해서 천잠의 vitellin은 glycolipoprotein으로 동정되었으며, 2개의 subunit도 당과 지질성분을 가지고 있었다. 4. 천잠 vitellin의 지질성분 중에는 가잠이나 Eri-참에 비하여 매우 다양한 종류들이 검출되며 특히 포화지방이라고 추정되는 물질도 있었다. 5. 천잠 vitellin의 단백질과 당성분은 7$0^{\circ}C$에 60분까지는 안전성을 보였으나, 같은 조건에서 지질성분은 검출되지 않았다. 6. 천잠 vitellin의 아미노산 성분 중 다른 곤충에서는 함량이 적은 glycine이 현격히 많았고 기타 성분의 함량은 가잠이나 Eri-잠과 같은 경향이었다. 7. 천잠의 vitellin은 직경이 14$\pm$0.8nm의 균일한 구형이었다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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• pp.102-102
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• 2001

Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.

• 한국응용곤충학회지
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• 제17권3호
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• pp.131-138
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• 1978

Cymbidium의 mild mosaic 병주로부터 각종 초본식물에 즙액접종되는 소구형 바이러스를 분리하여 다음과 같은 결과를 얻었다. 1. 본 바이러스를 즙액접종하였을 때 C. amaranticlor, C. quinoa, Cymbidium spp., Dianthus caryophyllus는 전신감염, C.ficifolium, Gomphrean globosa는 국부감염되었다. 2. 본 바이러스는 Myzus persicae로 전반되지 않고, 영양번식기관에 의하여 전반되었다. 3. 조즙액중의 불활성화한계는 내열성이 $90^{\circ}C$ (10분)이고 내희석성이 $10^{-6}$, 내보존성이 60일 $(20^{\circ}C)$이였다. 4. 본 바이러스는 C. amaranticolor병엽을 동결후, chlorform으로 청등하여 분획원심분리와 sucrose density gradient 원심분리법으로 순화하였다. 5. 순화시료의 자외선흡수곡선은 최고 261nm, 최저 243nm이고 260/280=1.72, 최고/최저=1.26의 핵단백에 의한 흡수곡선을 나타냈다. 침강계수는 $S_{20,w}=126$의 수치가 얻어졌다. 6. 本 바이러스의 항혈청은 침강반응혼합법에 의해 2,025배의 역가를 나타냈고, 한천내확산법에 의한 시험에서 CarMV와 혈청학적관계가 있는 것으로 나타났다. 7. 本 바이러스의 형태는 직경 약 28nm의 소구형(다면체) 입자이고, empty particles도 소수 관찰되었다. 5. 본 바이러스에 감염된 C. amaranticolor, C. ficifolium, Cymbidium spp.의 병엽초박절편을 전자현미경으로 관찰하였든 바, 각종 세포의 세포질, 액포 및 도관내에 소구형 입자가 산재 또는 집괴의 소재양식으로 확인되었다. 9. 이상의 결과를 종합해서 본 바이러스를 Cymbidium mild mosaic virus로 명명한다.