• Title/Summary/Keyword: Gene Identification

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Distribution and Identification of Halophilic Bacteria in Solar Salts Produced during Entire Manufacturing Process (천일염 생산공정별 미생물 분포 조사 및 호염미생물 동정)

  • Na, Jong-Min;Kang, Min-Seung;Kim, Jin-Hyo;Jin, Yong-Xie;Je, Jeong-Hwan;Kim, Jung-Bong;Cho, Young-Sook;Kim, Jae-Hyun;Kim, So-Young
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.133-139
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    • 2011
  • In this study, we determined the changes in microbial numbers in solar salts according to the manufacturing process and storage duration. The salt samples were harvested from salt farms in Shinan (area 2) and Yeonggwang (area 1). They were serially diluted ten-fold and then placed on 4 kinds of cultivable media (mannitol salt agar, eosin methylene blue, plate count agar, and trypticase soy agar). After incubation, we obtained 62 halophilic isolates from the salt samples. Coliform and general bacteria were not detected in all salt samples. By 16S rRNA sequencing analysis, we found 12 kinds of halophilic bacteria belonging to the genera Halobacillus, Halomonas, Bacillus, Idiomarina, Marinobacter, Pseudoalteromonas, Vibrio, Salinivibrio, Virgibacillus, Alteromonas, Staphylococcus and some un-known stains. In our study, we discovered two novel species that have a 16S rDNA sequence similarity below 97%.

Development of PCR assay for the detection of Brucella spp in bovine semen (종모우 정액중 Brucella균 신속 검출을 위한 PCR기법 개발)

  • Jung, Suck-chan;Jung, Byeong-yeal;Woo, Seong-ryong;Cho, Dong-hee;Kim, Jong-yeom;Kim, Woo-taek;Lee, Jung-mi;Park, Yong-ho;Baek, Byeong-kirl
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.345-352
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    • 1998
  • The diagnosis of brucellosis is currently based on serological and microbiological tests. However, the microbiological isolation and identification have several disadvantages such as time-consuming and laborious, and the serological methods have been reported to cross-react with antigens other than those from Brucella spp. To develop a sensitive and rapid diagnostic method for detection of Brucella species, the genus-specific primers were designed and synthesized from the sequence of gene encoding a 31kDa cell surface protein(BCSP) and a 36kDa outer membrane protein(OMPB) of B abortus. The amplified 711bp and 982bp DNA fragments were only visible in each species of Brucella by PCR method using the BCSP and OMPB primers, respectively. However, PCR product was not obtained with DNA from other Gram-negative bacteria. As little as 1pg of the B abortus genomic DNA could be detected by this PCR method. Using the PCR technique, semen samples from 185 bulls of Brucella-seronegative herds in Cheju island were examined for comparison of this PCR method with conventional methods in 1995. The semen samples from 5 bulls were positive by culture method and PCR, and one was positive and 5 were suspect by semen plasma agglutination test. However, the semen samples obtained from 177 bulls were negative by semen plasma agglutination, culture and PCR methods in 1996. The results of comparison tests suggested that PCR was a better test than agglutination test against semen of bulls. This study indicated that the PCR technique was a valuable for the diagnosis of bovine brucellosis, particulary in bull semens.

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Antibiotics Resistance and Molecular Analysis of Enterococcus Isolated from the Han-river in Korea (한강에서 분리한 장구균의 항생제 내성과 분자생물학적 분석)

  • Kwon, Oh-Min;Kim, Mal-Nam
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.116-124
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    • 2012
  • Identification was performed in March 2008 for the 76 Enterococcus strains isolated from the Han-river, which is used as water supply for Seoul citizens. The antibiotic susceptibility, antibiotic resistant structural analysis, trans-conjugation, pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) were also carried out for the isolated strains. Among the isolated strains, 25 strains were E. casseliflavus, 4 strains were E. faecalis and 1 strain was E. hirae. Investigation of antibiotic susceptibility indicated that 15 strains demonstrated tolerance against vancomycin, and that 11 strains of E. faecium and 4 strains of E. casseliflavus were VRE. The vanA gene detection of the VRE strains revealed that 6 E. faecium strains were vancomycin-resistant Enterococcus faecium (VREF) possessing vanA. Analyses of transposon Tn1546 structure containing vanA demonstrated that Km36 and Km37 belonged to Tn1 type, Km20 and Km38 was Tn2 type, and Km39 and Km40 was Tn3 type. PFGE disclosed that among the 6 VREF strains, Km36 and Km37 exhibited equivalent subtype, while the rest 4 strains showed subtypes different to each other. MLST for the 6 VREF strains disclosed that 3 strains were ST78, while the rest 3 strains were ST18, ST192 and ST230, respectively. All these clonal complexes were derived from CC17 which has been isolated from clinical sources. 4 strains belonged to CC78, while the rest 2 strains were CC18 and CC192, respectively.

Molecular and Morphologic Identification of Spirometra ranarum Found in the Stool of African Lion, Panthera leo in the Serengeti Plain of Tanzania

  • Eom, Keeseon S.;Park, Hansol;Lee, Dongmin;Choe, Seongjun;Kang, Yeseul;Bia, Mohammed Mebarek;Lee, Sang-Hwa;Keyyu, Julius;Fyumagwa, Robert;Jeon, Hyeong-Kyu
    • Parasites, Hosts and Diseases
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    • v.56 no.4
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    • pp.379-383
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    • 2018
  • The present study was performed with morphological and molecular analysis (cox1 and nad1 mitochondrial genes) to identify the proglottids of spirometrid tapeworm found in the stool of an African lion, Panthera leo, in the Serengeti plain of Tanzania. A strand of tapeworm strobila, about 75 cm in length, was obtained in the stool of a male African lion in the Serengeti National Park ($34^{\circ}$ 50' E, $02^{\circ}$ 30' S), Tanzania, in February 2012. The morphological features of the adult worm examined exhibited 3 uterine coils with a bow tie appearance and adopted a diagonal direction in the second turn. The posterior uterine coils are larger than terminal uterine ball and the feature of uteri are swirling rather than spirally coiling. The sequence difference between the Spirometra species (Tanzania origin) and S. erinaceieuropaei (GenBank no. KJ599680) was 9.4% while those of S. decipiens (GenBank no. KJ599679) differed by 2.1% in the cox1 and nad1 genes. Phylogenetic tree topologies generated using the 2 analytic methods were identical and presented high level of confidence values for the 3 major branches of the 3 Spirometra species in the cox1 gene. The morphological and molecular findings obtained in this study were nearly coincided with those of S. ranarum. Therefore, we can know for the first time that the African lion, Panthera leo, is to the definitive host of this tapeworm.

Diagnostic Value of Ceruloplasmin in the Diagnosis of Pediatric Wilson's Disease

  • Kim, Jung Ah;Kim, Hyun Jin;Cho, Jin Min;Oh, Seak Hee;Lee, Beom Hee;Kim, Gu-Hwan;Choi, Jin-Ho;Kim, Kyung Mo;Yoo, Han-Wook
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.18 no.3
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    • pp.187-192
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    • 2015
  • Purpose: Measurement of serum ceruloplasmin level is the first step in screening for Wilson's disease (WD). Despite the rarity of WD in the general population, ceruloplasmin levels are routinely measured through hepatitis screening in both adults and children. Herein, we evaluated the diagnostic value of ceruloplasmin for the diagnosis of WD among children with hepatitis. Methods: We retrospectively reviewed data on serum ceruloplasmin levels measured as a serologic marker for patients with hepatitis at Asan Medical Center (Seoul, Korea) between from January 2004 to November 2013. The diagnosis of WD was confirmed by the identification of pathogenic variants in the ATP7B gene. To determine the diagnostic accuracy of ceruloplasmin, receiver operation characteristic (ROC) curves were constructed and the area under curve (AUC) were calculated. Results: Measurements of serum ceruloplasmin were performed in 2,834 children who had hepatitis. Among these, 181 (6.4%) children were diagnosed with WD. The sensitivity, specificity, and accuracy of a ceruloplasmin level of <20 mg/dL in the discrimination of WD were 93.4%, 84.2%, and 84.8%, respectively. In this study, 418 (14.7%) false-positive cases and 12 (0.4%) false-negative cases were noted. Using a ROC curve, a ceruloplasmin level of ${\leq}16.6mg/dL$ showed the highest AUC value (0.956) with a sensitivity of 91.2%, a specificity of 94.9%, and an accuracy of 94.7%. Conclusion: The measurement of serum ceruloplasmin was frequently used for the screening of WD in children, despite a low positive rate. The diagnostic value of ceruloplasmin may be strengthened by adopting a new lower cut-off level.

Antimicrobial Susceptibility and Biochemical Characteristics of Streptococcus suis Isolated from Diseased Pigs in Gyeongbuk Province (경북지방 환돈에서 분리한 Streptococcus suis의 생화학적 성상 및 약제감수성)

  • Choi, Seong-Kyoon;Kim, Seong-Guk;Kim, Young-Hoan;Choi, Jeong-Hye;Jo, Min-Hee;Cho, Gil-Jae
    • Journal of Life Science
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    • v.20 no.10
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    • pp.1549-1555
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    • 2010
  • Streptococcus suis is a worldwide pathogen of a variety of porcine infection and has also been described as a pathogen for humans. We studied biochemical characteristics, antimicrobial susceptibility, and identification of polymerase chain reaction (PCR) of S. suis isolated from diseased pigs in Gyeongbuk province from 2004 to 2009. Sixty-one isolates were identified as S. suis by biochemical characteristics and PCR from 40 farms. The biochemical characteristics of S. suis isolates were production of VP-negative, hippurate, esculin, and arginine decarboxylase-positive, and fermentation of carbohydrate was variable lactose, trehalose, inulin, and raffinose, which was typeable 11 phenotype. In an antimicrobial susceptibility test, the majority of isolates were highly susceptible to amoxicillin/clavulanic acid, ampicillin, cephalothin, cefoperazone and florfenicol, while being highly resistant to streptomycin, kanamycin, amikacin, neomycin, erythromycin, clindamycin, and tetracycline. The isolates were divided into 11 phenotypes of biochemistry. By using PCR, the 16S-rRNA gene DNA fragment was detected at 304 bp from all of isolates. These results may provide the basic information needed to establish strategies for the prevention of S. suis infection in pigs.

Identification of QTLs for Some Agronomic Traits in Rice Using an Introgression Line from Oryza minuta

  • Rahman, Md Lutfor;Chu, Sang Ho;Choi, Min-Sun;Qiao, Yong Li;Jiang, Wenzhu;Piao, Rihua;Khanam, Sakina;Cho, Young-Il;Jeung, Ji-Ung;Jena, Kshirod K.;Koh, Hee-Jong
    • Molecules and Cells
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    • v.24 no.1
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    • pp.16-26
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    • 2007
  • Wild progenitor species provide potential gene sources for complex traits such as yield and multiple resistances to biotic and abiotic stresses, and thus are expected to contribute to sustainable food supplies. An introgression line 'IR71033-121-15' was derived from a wild species Oryza minuta (2n = 48, BBCC, Acc No. 101141) at IRRI. Introgression analysis using 530 SSR and STS markers revealed that at least 14 chromosomal segments distributed over 12 chromosomes had been introgressed from O. minuta. An $F_{2:3}$ population from the cross between IR71033 and Junambyeo (a Korean japonica cultivar) consisting of 146 lines was used for quantitative trait loci (QTL) analysis of 16 agronomic traits. A total of 36 single-locus QTLs (S-QTLs) and 45 digenic epistasis (E-QTLs) were identified. In spite of it's inferiority of O. minuta for most of the traits studied, its alleles contributed positively to 57% of the QTLs. The other QTLs originated from either parent, IR71033 or Junambyeo. QTLs for phenotypically correlated traits were mostly detected on introgressed segments. Fourteen QTLs corresponded to QTLs reported earlier, indicating that these QTLs are stable across genetic backgrounds. Twenty-two QTLs controlling yield and its components had not been detected in previous QTL studies. Of these, thirteen consisted of potentially novel alleles from O. minuta. QTLs from O. minuta introgression could be new sources of natural variation for the genetic improvement of rice.

Identification and gene expression profiling of chicken Pumilio family, Pum1 and Pum2

  • Lee, Jee-Young;Kim, Duk-Kyung;Zheng, Ying-Hui;Kim, Sun-Young;Kim, Hee-Bal;Lim, Jeong-Mook;Han, Jae-Yong
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2005.11a
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    • pp.64-65
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    • 2005
  • Members of the Pumilio are the RNA binding proteins acting as translational repressors and required for germ cell development and asymmetric division. We identified chicken Pum1 and Pum2 that are similar to mouse and human in highly conserved C-terminal RNA-binding domain and eight tandem repeats. The comparative sequence analysis of Pum1 and Pum2 from fly, chicken, mouse and human shows high degree of evolutionary conservation in the homology of the peptide sequence and the structure of PUM-HD (Pumilio homology domain) with similar spacing between adjacent Pum repeats. Also, structures of chicken Pum1 and Pum2 genes are almost identical to those of mouse and human. We revealed that the expression levels of Pum1 and Pum2 were the highest in hatched female gonad among various embryonic tissues, and Pum2 expressed highly in 12-day and hatched gonad by real-time RT-PCR. These results suggest that Pum1 and Pum2 might have an effect on the development of chicken gonad.

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Identification of Microorganisms from Eggs in Hypermarket in the Northern Gyeonggi Area (경기 북부 일부 지역 대형 마트 유통계란에 오염된 미생물의 분리)

  • Chun, Myoung-Sook;Hong, Seung-Hee
    • The Korean Journal of Food And Nutrition
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    • v.22 no.3
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    • pp.396-401
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    • 2009
  • Microorganisms or their toxins can be transferred to eggs and cause food poisoning in humans. Therefore, this study was conducted to detect microbial contamination of eggs and to identify microorganisms in any contaminated eggs. Four different brands of eggs were collected from hypermarkets in the northern Gyeonggi area. The total bacterial counts on the shells of the eggs varied greatly between brands. In addition, various bacterial species including Klebsiella pneumoniae, Pseudomonas mendocina, Alcaligenes xylosoxidans, Alcaligenes faecalis, and Enterobacter cloacae were identified on eggshells. Furthermore, mean of total bacterial counts of four brands was $3.4{\times}10^4 cfu/m{\ell}$ and E. coli was detected on the eggshell of one brand egg. However, Salmonella was not identified on all brands of collected eggs. We also demonstrated that the E. coli isolated from the eggshell was not pathogenic based on the absence of pathogen-specific gene expression patterns. Taken together, the result of this study indicate that strict quality control and improved distribution controls are required to decrease microbial contamination and improve human health.

A Thermostable Protease Produced from Bacillus sp. JE 375 Isolated from Korean Soil (한국의 토양으로부터 내열성 단백질 분해효소를 생산하는 Bacillus sp. JE 375의 선별)

  • Kim, Ji-Eun;Bai, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.38 no.3
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    • pp.419-426
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    • 2006
  • A thermophilic microorganism, strain JE 375, which produces a thermostable protease, was isolated from soil and compost in Korea. This gram-positive, rod-shaped, catalase positive, motility positive, and hemolysis ${\beta}$ containing organism was implicated in glucose fermentation, mannitol fermentation, xylose oxidation, aerobic activity and spore formation. The color of the colony was yellowish white. The temperature range for growth at pH 6.5 was between 55 and $70^{\circ}C$, with an optimum growth temperature of $65^{\circ}C$. This result confirmed the strain JE 375 as a thermophilic microorganism. The enzyme was produced aerobically at $65^{\circ}C$ during 20 hr in a medium (pH 6.5) containing 1% trypton. 1% maltose, 0.5% yeast extract and 1% NaCl. The 16S rDNA of strain JE 375 had 97.6% sequence similarity with the 16S rDNA of Bacillus caldoxyloyticus. On the basis of biochemical and physiological properties and phylogenetic analysis, we named the isolated strain as Bacillus sp. JE 375. The thermostable protease from Bacillus sp. JE 375 had been partially purified and characterized. The molecular weight of the enzyme was deduced from SDS-PAGE and gel chromatography as 55 kDa and its optimal temperature was $60^{\circ}C$. The enzyme showed its highest activity at pH 7.5 and was stable from pH 7.0 to 8.0.