• Title/Summary/Keyword: All-in-one gel

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One-step Separation of 30K Protein from the Silkworm Hemolymph by Anion-exchange Chromatography and Its Effect on the Proliferation of Human Cells (음이온교환 크로마토그래피를 이용한 누에체액 유래 30K 단백질의 정제와 정제된 단백질이 인간세포 배양 증식에 미치는 영향)

  • Shin Hyun-Chong;Joung Chan-Hi;Choi Yong-Soo;Lim Sang-Min;Han Kyuboem;Koo Yoon-Mo;Park Tai Hyun;Kim Dong-Il
    • KSBB Journal
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    • v.20 no.3
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    • pp.233-237
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    • 2005
  • In order to investigate the feasibility of 30K protein from silkworm (Bombyx mori) hemolymph (SH) on the proliferation of human cells, a simple separating procedure by anion-exchange chromatography system with Q-Sepharose fast flow gel was established. The 30K protein was eluted with an optimized condition of 0.16 M sodium chloride in 20 mM tris buffer (pH 9.0). The separated 30K protein at three concentrations of 0.04, 0.12, and 0.4 mg/ml was added to the culture medium with various human cells, such as chondrocytes, periosteum-derived cells, and MRC-5 cells, and their growth rates were measured. The cell growth rate at protein concentration of 0.4 mg/ml was always higher than that without 30K protein in all human cells tested, suggesting that the 30K protein has positive effect on the increase of the life span of human cells.

Estimation of Anti-proliferative Activity of Saccharin against Various Cancer Cell Lines and MSCs (다양한 암세포 주와 MSCs에 대한 Saccharin의 항증식성 평가)

  • Choi, Jeong Su;Park, Sang Yong;Yang, Man Gil;Lee, Dong Beom;Lee, Tae Bok;Heo, Ji Hye;Lee, Min Woo;Kim, Suhng Wook
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.169-175
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    • 2016
  • Saccharin (o-benzoic sulfimide) is the first artificial and non-caloric sweetener that was first synthesized in 1879. In this study, we examined the biological activity of saccharin against various human cancer cell lines and human bone marrow-derived mesenchymal stem cells. A viability assay based on the conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed to test for the cytotoxicity of saccharin about the four human cancer cell lines (H460, H157, A549 and SKOV3), one murine cancer cellline (Raw264.7), and MSCs. In order to find the differentially expressed gene in saccharin-treated MSCs against untreated MSCs, we performed annealing control primer (ACP)-based differential display reverse transcriptionp-olymerase chain reaction (DDRT-PCR). All tested cells were treated with saccharin at various concentrations (0.0, 4.8, 7.2, 9.6, 12.0, 14.4 mg/mL) for 48 hr. The number of metabolically active cancer cells decreased when treated with the saccharin at various concentrations for 48 hr as compared with the untreated cells. The decrease in cell survival was more evident with increasing concentrations of saccharin. Moreover, novel candidate genes, which were differentially expressed in MSCs in response to saccharin, were identified in 16 bands on 2% agarose gel. This revealed 16-7 up-regulated and 9 down-regulated-differentially expressed genes indicated by arrows. One of these candidate genes was a FK506-binding protein gene. The functional roles of FK506 binding proteins, with respect to the activities of stem cell proliferation, were not characterized. Further studies are required to get a better understanding of FK506-binding proteins in its roles in increasing stem cell proliferative activities from using saccharin.

Difference of Classification, Growth and Herbicidal Tolerance in Collected Weedy Rice(Oryza sativa) (수집(蒐集) 잡초성(雜草性)벼(Oryza sativa)의 분류(分類), 생장(生長) 및 제초제(除草劑) 내성차이(耐性差異))

  • Kuk, Y.I.;Guh, J.O.;Chon, S.U.
    • Korean Journal of Weed Science
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    • v.17 no.1
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    • pp.31-43
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    • 1997
  • This study was carried out to investigate classfication of weedy rice (Oryza sativa) based on isozymes esterase and peroxidase, growth and developmental difference of weedy rices and rices grown under dry and water condition, and weedy rice control and tolerant difference of weedy rices in various herbicides using weedy rices collected from thirteen strains of Chonnam, one Chonbuk, two Kyeongki and two rice cultivars. 1. The collected weedy rices were classified into three groups based on isozyme esterase and peroxidase using polyacrylamide gel electrophoresis(PAGE) method. The classified groups were not same each other. 2. Plant height was taller in collected weedy rices than rice cultivars at 18 days after seeding under dry and water conditions, but number of leaves, shoot fresh weight, root fresh weight and root length were not significantly different between collected weedy rices and rice cultivars. In addition, growths of collected weedy rices were greater in dry- than water-condition. 3. After thiobencarb(S-4-chlorobenzyl diethythiocarbamate), molinate(S-ethyl hexahydro-1H-azepine-1-carbothioate) and oxadiazon(5-tert-butyl-3(2,4-dichloro-5-isopropoxyphenyl)-1,3,4-oxadiazol-2-one) were applied at 6 days before seeding, the weedy rices controlled 100% by thiobencarb at 2.1kg ai/ha and 024kg ai/ha oxadiazon treatment but controlled 26% to 67% by molinate at 6.5kg ai/ha. Rice due to the herbicides was injured severely(25% to 100%) in flood condition at time of rice seeding after oxadiazon at 0.48kg ai/ha and 2.1kg ai/ha thiobencarb application, except for molinate which injured rice slightly(4% to 13%) in drain condition. The collected weedy rices to all experimented herbicides showed slight intraspecific variations. The intraspecific variations of weedy rices decreased in the order of thiobencarb>molinate>oxadiazon.

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Preparation of Soluble Dietary Fiber from Oak Wood (Quercus Mongolica) and Its Physiological Function in Rat Fed High Cholesterol Diets (참나무 (Quercus Mongolica)로부터 수용성 식이섬유소의 제조 및 기능성 검증)

  • 채영미;임부국;이종윤;김영희;이순재
    • Journal of Nutrition and Health
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    • v.36 no.1
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    • pp.9-17
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    • 2003
  • The preparation method of a soluble dietary fiber from oak wood (Quercus mongolica) and the effect of the soluble dietary fiber on physiological function in rat fed high cholesterol diets was investigated. The best condition for steam explosion method was 25 kgf/㎤ pressure for 6 min. The exploded samples were delignified by the filtration treatment with 1% NaOH for several times, which is the best condition. The enzymatic hydrolysis of Cellusoft cellulase was more effective than Onozuka R-10 cellulase. The manufactured soluble dietary fiber was assayed using gel permeation chromatography (GPC) and it was dissolved in water. Average molecular weight distribution of manufactured soluble dietary fiber was about 348-1,200 and it was assumed the oligomer form fraction. In order to compare the manufactured soluble dietary fiber with commercial soluble dietary fiber (pectin) on the physiological function, Sprague-Dawley male rats weighing 100$\pm$10 g were randomly assigned to one normal diet and five high cholesterol diet containing 1% cholesterol. The high cholesterol diet groups were classified to fiber free diet (FF group), 5% pectin (5P group), 10% pectin (l0P group), 5% manufactured soluble dietary fiber (5M group) and 10% manufactured soluble dietary fiber (10M group). Body weight gains in all soluble dietary fiber groups were lower than FF group. Food intakes were increased in all soluble dietary fiber groups than that of FF group. Food efficiency ratio (FER) was significantly decreased in all soluble dietary fiber groups than that of the FF group, and it was especially was highest in 10% supplemented soluble dietary fiber group. The weight of liver of the soluble dietary fiber supplemented groups were lower than those of the FF group, but weights of cecum and small intestine of all supplemented soluble dietary fiber groups were significantly increased, compared with that of FF group. The weights and water contents in feces were significantly increased by the soluble dietary fiber. The activity of the glutamic oxaloacetic transaminase in soluble dietary fiber groups were significantly decreased than those of FF group. The hepatic glutathione S-transferase activity in all soluble dietary fiber supplemented groups were higher than that of FF group. The physiological effects of the manufactured soluble dietary fiber are the same as the commercial soluble dietary fiber (pectin). The preparation method of the soluble dietary fiber from the oak chips suited to its purpose. (Korean J Nutrition 36(1) : 9~17, 2003)

Powdery Mildew Resistance Phenotype Test & Genotype Test in C. moschata

  • Jong-Gyu Park
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.290-290
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    • 2022
  • Powdery mildew is known to be one of the serious diseases in C. moschata cultivation. Plants infected with powdery mildew cause damage to cultivation areas such as occurrence of deformity fruit and decrease in quantity. also, it has been reported that many farms have difficulties in controlling powdery mildew due to the outbreak under various conditions throughout the year. Therefore, this study intends to perform a phenotype test and a genotype test for C. moschata 60 lines grown in Jenong S&T. Podospareaxanthii, known as a pathogen that causes powder mildew disease in pumpkins in Korea, was collected and used as an inoculation source, phenotype test was performed by examining the infection area rate(%) of powdery mildew disease that occurred in leaves 25 days after inoculation. It was determined that 0% of the infection area rate was in the first stage, 1 to 5% in the second stage, 6 to 15% in the third stage, 16 to 30% in the fourth stage, and 31% or more in the fifth stage, The first and second stages were judged as resistance, the third as moderate resistance, and the fourth and fifth stages as sensitivity. As a result of the phenotype test, it was confirmed that the resistance was 21 points, moderate resistance was 14 points, and sensitivity was 25 points. After searching for the genes related to powdery mildew resistance resistance, pm-0, CmbHLH87, and LOC111453072, 21 points of resistance and 9 points of moderate resistance identified through phenotype tests were identified through gel electrophoresis after polymerase chain reaction(PCR) using 5 primers related to 3 genes. As a result of genotype testing of a total 30 points, the CmbHLH87 and LOC111453072 gene were found to be resistant bands in all points, PMR1 was identified as 20 points for resistance, 4 points for moderate resistance, and 6 points for sensitivity, PMR2 was not identified in the entire band, and PMR5 was identified as 18 point for resistance, 3 points for moderate resistance, and 9 points for sensitivity. As a result, when comparing the phenotype test results and genotype test results, CmbHLH87 and LOC111453072 genes was 100% consistent in resistance and moderate resistance, PMR1 was 95.2% in resistance, 44.4% in moderate resistance, and PMR5 was 90% in resistance and 33.3% in moderate resistance, PMR2 was not consistent in resistance and moderate resistance. Therefore, it is expected that more accurate PMR test will be possible by using molecular markers(PMR1, PMR5) and by developing CmbHLH87 and LOC111453072 gene-related molecular markers.

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Study of Viral Effects of the Mycovirus (LeV) and Virus-Free Commercial Line in the Edible Mushroom Lentinula edodes

  • Kim, Jung-Mi;Song, Ha-Yeon;Yun, Suk-Hyun;Lee, Hyun-Suk;Ko, Han-Kyu;Kim, Dae-Hyuk
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.37-37
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    • 2015
  • dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

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Molecular Characteristics of Extended-Spectrum β-Lactamase Genes in Clinical Isolates of Escherichia coli and Klebsiella pneumoniae (임상검체에서 분리된 Escherichia coli와 Klebsiella pneumoniae의 Extended-Spectrum β-Lactamase 유전자형 및 분자유전학적 특성)

  • Chung, Kyung-Suk
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.1
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    • pp.26-33
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    • 2006
  • Recently, the rapid increase in extended-spectrum ${\beta}$-lactamase (ESBL) producing clinical isolates has become a serious problem. In this study, the epidemiologic features and molecular characteristics of ESBL among clinical isolates of Escherichia coli and Klebsiella pneumoniae, antibiotic susceptibility testing, genotype of the ESBL and patterns of chromosomal DNA from PFGE (pulsed field gel electrophoresis) were observed. A total of 53 ESBL-producing clinical isolates (30 of E. coli and 23 of Klebsiella pneumoniae) were collected from two university hospitals in the period of June to July in 2002 and 2003 respectively. The antibiotic resistance frequency of those 53 strains was tested by the disk agar diffusion method with the result that all the strains were resistant to cephalothin. To other antibiotics, the resistance rates of E. coli (30 isolates) were in order of ceftazidime (90.0%), cefotaxime and aztreonam (respectively 83.3%). Also, the resistance rates of K. pneumoniae (23 isolates) were in order of aztreonam (78.3%), ceftazidime (73.9%) and cefotaxime (65.3%). Also the sensitivity of ceftazidime-clavulanic acid were 100% in E. coli and 95.7% in K. pneumoniae. And the sensitivity of cefotaxime-clavulanic acid was 96.7% in E. coli and 91.3% in K. pneumoniae. The types of the ESBL genes were determined by using polymerase chain reaction (PCR). Among the 30 isolates of ESBL-producing E. coli, 6 (20.0%) have SHV only, 5 (16.7%) have TEM only and, 18 (60.0%) have both of TEM and SHV. Among the 23 isolates of ESBL-producing K. pneumoniae, 7 (30.4%) have SHV only, 2 (8.7%) have TEM only, and 14 (60.9%) have both of TEM and SHV. These results show that 52 strains, with only one exception, were confirmed as either TEM or SHV. The patterns of Xba I-digested chromosomal DNA of ESBL-producing E. coli and K. pneumoniae isolates were analyzed by PFGE. PFGE patterns of E. coli and K. pneumoniae were multiclonal, but many strains were grouped into a few types. Therefore, it seems that there were clonal outbreaks or possible horizontal spread. In conclusion, the TEM and SHV ${\beta}$-lactamase are most widely spread in E. coli and K. pneumoniae in Korea. As these types are usually carried by plasmids, the spread of these ${\beta}$-lactamase genes could compromise the future usefulness of third generation cephalosporins for the treatment of infections caused by E. coli and K. pneumoniae.

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Effects of cultivation ages and modes on microbial diversity in the rhizosphere soil of Panax ginseng

  • Xiao, Chunping;Yang, Limin;Zhang, Lianxue;Liu, Cuijing;Han, Mei
    • Journal of Ginseng Research
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    • v.40 no.1
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    • pp.28-37
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    • 2016
  • Background: Panax ginseng cannot be cultivated on the same land consecutively for an extended period, and the underlying mechanism regarding microorganisms is still being explored. Methods: Polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) and BIO-LOG methods were used to evaluate the microbial genetic and functional diversity associated with the P. ginseng rhizosphere soil in various cultivation ages and modes. Results: The analysis of microbial diversity using PCR-DGGE showed that microbial communities were significantly variable in composition, of which six bacterial phyla and seven fungal classes were detected in P. ginseng soil. Among them, Proteobacteria and Hypocreales dominated. Fusarium oxysporum, a soilborne pathogen, was found in all P. ginseng soil samples except R0. The results from functional diversity suggested that the microbial metabolic diversity of fallow soil abandoned in 2003was the maximum and transplanted soil was higher than direct-seeding soil and the forest soil uncultivated P. ginseng, whereas the increase in cultivation ages in the same mode led to decreases in microbial diversity in P. ginseng soil. Carbohydrates, amino acids, and polymers were the main carbon sources utilized. Furthermore, the microbial diversity index and multivariate comparisons indicated that the augmentation of P. ginseng cultivation ages resulted in decreased bacterial diversity and increased fungal diversity, whereas microbial diversity was improved strikingly in transplanted soil and fallow soil abandoned for at least one decade. Conclusion: The key factors for discontinuous P. ginseng cultivation were the lack of balance in rhizosphere microbial communities and the outbreak of soilborne diseases caused by the accumulation of its root exudates.

Case Report for a Large-Scale Food Poisoning Outbreak that Occurred in a Group Food Service Center in Chungnam, Korea (2020년 충남지역 집단급식소에서 발생한 대형 식중독의 사례 보고)

  • Lee, Hyunah;Kim, Junyoung;Nam, Hae-Sung;Choi, Jihye;Lee, Dayeon;Park, Seongmin;Lim, Ji-Ae;Cheon, Younghee;Choi, Jinha;Park, Junhyuk
    • Journal of Environmental Health Sciences
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    • v.46 no.5
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    • pp.525-531
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    • 2020
  • Objectives: This study was performed to identify the epidemiological features of a food poisoning outbreak in a company cafeteria located in Chungcheongnam-do Province, Korea in June of 2020 and to suggest preventive measures for a similar incidence. Methods: A total of 84 patients with acute gastroenteritis were examined. Environmental samples were obtained from 16 food handlers, six food utensils, 135 preserved foods served over three days and nine menus, and six drinking water samples. These are analyzed to detect viruses and bacteria. Results: Ninety-four out of the 402 people who were served meals (23.4%) predominantly showed symptoms of diarrhea, and the number was over 3 times. Among the 84 patients under investigation, 17 cases (20.2%) were positive for Enteropathogenic E. coli (EPEC) and 18 cases were positive for Clostridium (C.) perfringens (21.4%). Based on the investigation, it was concluded that the main pathogens were EPEC and C. perfringens. For EPEC, it was detected in three of the food service employees and in the preserved food and curry rice. The results of pulsed field gel electrophoresis indicate that all EPEC cases are closely related except for one food service employee. Assuming that isolated EPEC originated from the preserved food, the incubation period is about 25 hours. The origin of the C. perfringens was not determined as it was not detected in the food service employees or environmental samples. Conclusions: This case suggests that food provided in group food service centers must be thoroughly managed. In addition, identifying the pathogens in preserved food is very important for tracing the causes of food poisoning, so food must be preserved in an appropriate condition. To prevent similar food poisoning cases, analyzing cases based on epidemiological investigation and sharing the results is needed.

Structural Analysis of Class I Integron Gene Cassette and Assessment of Genetic Relationships by PFGE of Salmonella enterica Serovar Typhimurium Isolated in Gyeongbuk Area (경북지역에서 분리된 Salmonella enterica Serovar Typhimurium의 Class I Integron Gene Cassette 특성과 PEGE 유형분석)

  • Sohn, Chang-Kyu;Lee, Jung-A;Lee, Do-Young;Hun, Wan;Jung, Jung-Kyo
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.12-18
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    • 2006
  • Thirty five Salmonella enterica serovar Typhimurium strains were isolated from diarrheic patients and pigs in Gyeongbuk area from 2003 to 2004. All 35 strains (17 strains from diarrheic patients and 18 from pigs) were resistant to more than one drug and most of strains isolated from pigs were resistant to ampicillin, ohloram-phenicol, streptomycin, sulfamethoxazole-trimethoprime, tetracyclin and nalidixic acid. Each isolate was also screened or the presence of class I, II and III integron gene cassettes. Among 35 strains,3 out of 17 strains isolated from diarrheic patients, carried dhfrX-orfF-aadA2 integron gene cassette and among 18 strains isolated from diseased pigs, 11 strains carried dhfrX-orfF-aadA2 integron gene cassette and 1 strain carried aadA2 integron only. But any class II and class II integron gene cassette were not detected in 35 strains. Thirty five strains were divided by five pulsotypes. Thirty one strains out of thirty five were pulsotype A. Among the remaining 4 strains, one each strain belonged to pulsotype B, C, D and pulsotype E. This data of pulsotypes showed that the widespread of pulsotype A, Salmonella enterica serovar Typhimurium in human and pigs in Gyeongbuk area may have been caused by the dissemination of a few epidemic strains in this area. Thirteen strains contain dhfrX-orfF-aadA2 integron gene cassette showed pulsotype A and one strain contains dhfrX-orfF-aadA2 integron gene cassette showed pulsotype B. One strain contains aadA2 integron showed pulsotype E. But fifteen strains do not contain any integron showed pulsotype A.