• Title/Summary/Keyword: New strains

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Pulsed-Field Gel Electrophoresis and Monoclonal Antibody Analysis of Leptospira interrogans Isolated in Korea (국내 분리 렙토스피라균의 단클론 항체 및 Genomic DNA의 Pulsed-Field Gel Electrophoresis 분석)

  • 조민기;기선호;김형준;김윤원;장우현;오희복
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.197-204
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    • 1999
  • A total of 22 Leptospiua inlermgans field isolates from the ~ a t s captured in 5 provinces of Korea in 1996, and 6 antigenically closely related relerence serovars of lai, yeonchon, birkini. gem, mwogolo. and canicola were analysed. When the antigenic characteristics were analysed by reactivity with 7 monoclonal antibodies prepared with sh.ains belongng to serogroup Icterohaemorrhagiae. all 22 isolates showed the same reaction pattern with that of serovar lai. Large restriction fragment patterns obtained after cleavage of geno~nic DNAs with infrequently cuttimg restriction enzymes were analyzed by pulsed-field pel electrophoresis(PFGE). Identification of leptospira strains by PFGE with Nor I, Asc I or Iise I digests correlated with their antigenically typed serovars, silh a few exceptions. PFGE of isolates, except for JR89, digested wjth Nor I showed identical pattern w~th serovar lai, showing 13 Cragments between 940 kb and 63 kb. When PFGE pallerns of JR89 were compared with those of serovar lai, Not I digest showed additional two hands of 1000 kb and 460 kb, while Asc I digest showed 650 kb fragment and Fse I digest did not show the fragment of 280 kb. Whereas serovar yeonchon. which was isolated in Korea and identified as a new serovar previously. could be differentiated from serovar lai in antigenic reactivities with monoclonal antibodres. it showed the similar PFGE pattern with serovar lai includin~ reference and field isolates. It was suggested that Korean leptospiral field isolates are closely related in DNA level.

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A study of serotyping of Streptococcus pneumoniae by multibead assay (다중구슬 분석법에 의한 폐구균 혈청형 결정 연구)

  • Cho, Ky Young;Lee, Jung Ah;Cho, Sung Eun;Kim, Nam Hee;Lee, Jin A;Hong, Ki Sook;Lee, Hoan Jong;Kim, Kyung Hyo
    • Clinical and Experimental Pediatrics
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    • v.50 no.2
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    • pp.151-156
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    • 2007
  • Purpose : Streptococcus pneumoniae is a major etiologic agent for pneumonia, meningitis, otitis media, and sepsis among young children. Multi-drug resistant strains have raised great concern worldwide, thus the importance of prevention with vaccines has been emphasized. However, vaccines may force the appearance of pneumococcal infections by nonvaccine serotypes. Thus, distribution of pneumococcal serotypes should be monitored to estimate vaccine efficacy. We used a new and efficient multibead assay in determining pnemococcal serotypes. Methods : From January to February 2005, 643 children were recruited from ten day care centers to isolate pneumococci from their oropharynx. Pneumococcal serotyping was performed on 62 pneumococcal isolates from 60 children by multibead assay. This immunoassay required two sets of latex particles coated with pneumococcal polysaccharides and serotype-specific antibodies. Twenty four newly developed monoclonal antibodies specific for common serotypes and a pool of polyclonal rabbit sera for some of the less common serotypes were used. Results : The most prevalent pneumococcal serotypes were serotype 6A, 19A, 19F, 23F, and 11A/D/F which accounted more than 50 precent of all the 62 pneumococcal isolates. We found that multibead assay can be performed very rapidly and objectively. Conclusion : This multibead immunoassay was very useful in serotyping clinical isolates of S. pneumoniae because it was simple, reliable and fast.

Functional analysis of seaR protein identified from Saccharopolyspora erythraea (희소방선균의 seaR 단백질 발현을 통한 기능 분석)

  • Ryu, Jae Ki;Kwon, Pil-Seung;Lee, Hyeong Seon
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.39-47
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    • 2015
  • Secondary metabolism in actinomycetes has been known to be controlled by a small molecule, ${\gamma}$-butyrolactone autoregulator, the binding of which to each corresponding receptor leads to the regulation of the transcriptional expression of the secondary metabolites. We expected that expression of an autoregulator receptor or a pleiotropic regulator in a non-host was to be gained insight of effective production of new metabolic materials. In order to study the function of the receptor protein (seaR), which is isolated from Saccharopolyspora erythraea, we introduced the seaR gene to Streptomyces coelicolor A3(2) as host strains. An effective transformation procedure for S. coelicolor A3(2) was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a ${\varphi}C31$-derived integration vector, pSET152, which contained int, oriT, attP and $ermEp^*$ (erythromycin promotor). Therefore, the pEV615 was introduced into S. coelicolor A3(2) by conjugation and integrated at the attB locus in the chromosome of the recipients by the ${\varphi}C31$ integrase (int) function. Exconjugant of S. coelicolor A3(2) containing the seaR gene was confirmed by PCR and transcriptional expression of the seaR gene in the transformant was analyzed by RT-PCR. In case of S. coelicolor A3(2), a phenotype microarray was used to analyze the phenotype of transformant compared with wild type by seaR expression. After that, in order to confirm the accuracy of the results obtained from the phenotype microarray, an antimicrobial susceptibility test was carried out. This test indicated that sensitivity of the transformant was higher than wild type in tetracycline case. These results indicated that some biosynthesis genes or resistance genes for tetracycline biosynthesis in transformant might be repressed by seaR expression. Therefore, subsequent experiments, analysis of transcriptional pattern of genes for tetracycline production or resistance, are needed to confirm whether biosynthesis genes or resistance genes for tetracycline are repressed or not.

Paenibacillus kimchicus sp. nov., an antimicrobial bacterium isolated from Kimchi (김치로부터 분리된 항균 활성 세균 Paenibacillus kimchicus sp. nov.)

  • Park, A-rum;Oh, Ji-Sung;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.319-326
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    • 2016
  • An antimicrobial bacterium to pathogenic microorganisms, strain $W5-1^T$ was isolated from Korean fermented-food Kimchi. The isolate was Gram-staining-variable, strictly aerobic, rod-shaped, endospore-forming, and motile with peritrichous flagella. It grew at $15-40^{\circ}C$, at pH 6.0-10.0, and in the presence of 0-4% NaCl. Strain $W5-1^T$ could hydrolyze esculin and xylan, and assimilate $\small{D}$-mannose, but not $\small{D}$-mannitol. Strain $W5-1^T$ showed antimicrobial activity against Listeria monocytogens, Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella typhi. The G+C content of the DNA of strains $W5-1^T$ was 52.6 mol%. The predominant respiratory quinone was menaquinone-7 (MK-7) and the major cellular fatty acids were $C_{16:0}$, antieiso-$C_{15:0}$, $C_{18:0}$, and $C_{12:0}$. The strain contained meso-diaminopimelic acid in cell-wall peptidoglycan. On the basis of 16S rRNA gene sequence and phylogenetic analysis, the strain W5-1 was shown to belong to the family Paenibacillaceae and was most closely related to Paenibacillus pinihumi $S23^T$ (98.4% similarity) and Paenibacillus tarimensis $SA-7-6^T$ (96.4%). The DNA-DNA relatedness between the isolate and Paenibacillus pinihumi $S23^T$ was 8.5%, indicating that strain $W5-1^T$ represented a species in the genus Paenibacillus. On the basis of the evidence from this polyphasic study, it is proposed that strain $W5-1^T$ is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus kimchicus sp. nov. is proposed. The type strain is $W5-1^T$ (=KACC $15046^T$ = $LMG 25970^T$).

Identification of a new marine bacterium Ruegeria sp. 50C-3 isolated from seawater of Uljin in Korea and production of thermostable enzymes (대한민국 울진 연안 해양에서 분리한 해양 미생물 Ruegeria sp. 50C-3의 동정 및 내열성 효소 생산)

  • Chi, Won-Jae;Kim, Jong-Hee;Park, Jae-Seon;Hong, Soon-Kwang
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.344-351
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    • 2016
  • A marine bacterium, designated as strain 50C-3, was isolated from a seawater sample collected from the East Sea of South Korea. The strain is a Gram-negative, aerobic, yellow colored polar-flagellated bacterium that grows at $20-50^{\circ}C$ and pH 5.5-8.5. Optimal growth occurred at $40-50^{\circ}C$, at pH 6.5-7.5, and in the presence of 2% (w/v) NaCl. Based on 16S rRNA gene sequence similarity, the isolate was considered to represent a member of the genus Ruegeria. The result of this analysis showed that strain 50C-3 shared 99.4% and 96.98% sequence similarity with Ruegeria intermedia CC-GIMAT-$2^T$ and Ruegeria lacuscaerulensis ITI-$1157^T$, respectively. Furthermore, strain 50C-3 showed clear differences from related strains in terms of several characteristics such as motility, carbon utilization, enzyme production, etc. The DNA G+C content was 66.7 mol%. Chemotaxonomic analysis indicated ubiquinone-10 (Q-10) as the predominant respiratory quinone. Based on phenotypic, chemotaxonomic, and phylogenetic characteristics, the isolate represents a novel variant of the Ruegeria intermedia CC-GIMAT-$2^T$, for which we named Ruegeria sp. 50C-3 (KCTC23890=DSM25519). Strain 50C-3 did not produce cellulase and agarase, but produced alkaline phosphatase, ${\alpha}$-galactosidase, and ${\beta}$-galactosidase. The three enzymes showed stable activities even at $50^{\circ}C$ and thus regarded as thermostable enzymes. Especially, the ${\beta}$-galactosidase activity enhanced by 1.9 times at $50^{\circ}C$ than that at $37^{\circ}C$, which may be very useful for industrial application.

Agronomic Traits of Soybean Breeding Lines with Low Stachyose and Raffinose Contents (Stachyose 및 Raffinose 저함량 콩 선발계통의 농업적 형질)

  • Ha, Do Su;Moon, Jin Young;Choi, Sang Woo;Shim, Sang In;Kim, Min Chul;Chung, Jong Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.62 no.2
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    • pp.143-148
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    • 2017
  • Soybean [Glycine max (L.) Merr.] seed is an important dietary source of protein, oil, carbohydrates, isoflavones, and other nutrients for humans and animals. Raffinose and stachyose are the main antinutritional factors in soybean seed. They are carbohydrates belonging to the raffinose family of oligosaccharides, which are not readily digested in humans and cause flatulence or diarrhea. The genetic reduction of the raffinose and stachyose contents in mature soybean seeds will improve the nutritional value of soybean. The objective of this research was to evaluate agronomic traits with 10 $F_6$ strains selected from breeding populations derived from a cross among seven parents. The contents of raffinose and stachyose in mature seeds were detected by high-performance liquid chromatography. Agronomic traits such as flower color, flowering date, harvesting date, lodging, plant height, seed coat color, hilum color, 100 seed weight, and yield were evaluated. Ten intermediate parents showed low raffinose and stachyose contents. The intermediate parent 883-1 had a small seed size, six intermediate parents (15A1, 15D1, RS-5, RS-33, RS-64, and RS-70) had a medium seed size, and two intermediate parents (14G20 and RS-21) had a large seed size. The intermediate parent RS-21 had a black seed coat and a green cotyledon. Four intermediate parents (883-1, 14G20, RS-5, and RS-21) had elite agronomic traits. The new intermediate parents developed through this study will be used to develop improved soybean cultivars with low contents of raffinose and stachyose.

Studies on the Production of Alcohol Fermented Milk (알코올 발효유(醱酵乳)의 제조(製造)에 대(對)하여)

  • Yoo, Jin-Young;Kang, Tong-Sam;Min, Byong-Yong
    • Korean Journal of Food Science and Technology
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    • v.10 no.3
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    • pp.337-343
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    • 1978
  • In order to develope a new kind of fermented milk, basic studies on several lactic acid bacteria and yeasts were conducted, 8 kinds of alcohol fermented milk were manufactured and sensory evaluation was undertaken. The results obtained are summarized as follows: 1. Four kinds of lactic acid bacteria were isolated, among which Y-2 strain was strongest in acid productivity and it was elucidated that acid productivity of all strains was stornger in synthetic medium than in milk medium. 2. The pH in milk medium inoculated with Y-2 strain and incubated at $30^{\circ}C$ for 24 hours was dropped from 5.8 to 3.8 and fluctuation in amino nitrogen content was found during incubation. 3. The pH in milk medium inoculated with K. fragilis and incubated at $30^{\circ}C$ for 7 days was dropped from 6.2 to 5.2 and amino nitrogen content was in the range of $0.12{\sim}0.27mg/ml$. Alcohol productivity of K. fragilis was stronger than E-2 and E-4 strain but no difference in alcohol productivity was found between milk medium and synthetic medium. 4. The repression in growth and acid productivity of lactic acid bacteria was recongnized if inoculated after inoculating yeast firstly. 5. Alcohol productivity was increased rapidly at the end of acid production of lactic acid bacteria if lactic acid bacteria if lactic acid bacteria and yeast were inoculated simultaneously. 6. Sensory evaluation showed that the product that alcohol content and acidity were 1% and 0.8% respectively had the best palatability(p<0.01). 7. Chemical composition of final product was similar to that of milk koumiss in ash, protein and moisture content.

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Characteristics and pedigree selection of a shortened cultivation period strain in Lepista nuda (재배기간이 짧은 민자주방망이버섯 우량계통 선발 및 특성)

  • Jeon, Jong-Ock;Lee, Kwan-Woo;Lee, Kyoung-Jun;Kim, Min-Ja;Kim, In-Jae;Kim, Young-Ho
    • Journal of Mushroom
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    • v.18 no.4
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    • pp.331-338
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    • 2020
  • This study was conducted to cultivate new Lepista nuda varieties with shorter cultivation period and better fruiting body compared to that of wild strains, for mass production and commercial application. Eighteen genetic resources of L. nuda were collected and grown in boxes using rice straw-fermented growth medium. Four lines with fruiting bodies were formed and selected as cross-breeding lines. Although 657 combinations were crossed through monospore crossing, only 17 combinations were bred between the 'CBMLN-19' line and the 'CBMLN-30' line. Among them, 8 lines with fast mycelial growth and high density were selected. After inoculating the rice straw-fermented growth medium with 14 genetic resources and 8 cross-breeding lines, their incubation period was investigated. Six of the cross-breeding lines completed their incubation in 20 days, while 7 of the 14 genetic resources took more than 40 days to complete their incubation, reducing the incubation period by more than 20 days in most cross-breeding lines. After the incubations were completed, the clay loam soil was covered with for post-cultivation, and when the mycelial cultivation was complete, the formation of fruiting bodies was induced after scraping the mycelial bodies under these environmental conditions: 14℃, 95% relative humidity or higher, and 1,500 to 2,000 ppm CO2 concentration. The temperature was reduced to 6℃ at night, resulting in a low temperature shock. Thus, 4 lines of fruiting bodies occurred from two genetic resources 'CBMLN-31' and 'CBMLN-44' and two cross-bred lines 'CBMLN-96' and 'CBMLN-103'. After inoculation, the longest period for fruiting bodies to occur was 100 days for the control:, the genetic resource 'CBMLN-31', and the shortest period (45 days) was observed for the cross-breeding line 'CBMLN-103'. The result of the investigation of the fruiting body characteristics shows that the cross-bred line 'CBMLN-103' showed a small form with 1.9 g of individual weight and 123validstipes per box, which was the highest incidence among the four lines. Another cross-bred line, 'CBMLN-96', had an individual weight of 5.5 g, which is larger than that of 'CBMLN-103'; however, the number of valid stipes per box was 30 less than that of 'CBMLN-103'. Quantity analysis showed that the control, 'CBMLN-31', had the highest quantity of 783 g per box, followed by the cross-bred line, 'CBMLN-96' with 165 g per box, and then the 'CBMLN-103' with 232 g. The quantity of the two crossbred lines was lower than that of the control 'CBMLN-31'; however, the amount of fruiting bodies was higher, and the cultivation period was shortened by 32 to 33 days. Therefore, these two lines would be selected as superior lines.

Development and Evaluation of Multiplex PCR for the Detection of Carbapenemase-Producing Enterobacteriaceae (카바페넴분해효소 생성 장내세균 검출을 위한 Multiplex PCR의 개발 및 평가)

  • Kim, Si Hyun;Bae, Il Kwon;Kim, Na Young;Song, Sae Am;Kim, Sunjoo;Jeong, Joseph;Shin, Jeong Hwan
    • Annals of Clinical Microbiology
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    • v.22 no.1
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    • pp.9-13
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    • 2019
  • Background: The isolation of carbapenemase-producing Enterobacteriaceae (CPE) has become increasingly common. Continuous surveillance for these organisms is essential because their infections are closely related to outbreaks of illness and are associated with high mortality rates. The aim of this study was to develop and evaluate multiplex PCR as a means of detecting several important CPE genes simultaneously. Methods: We aimed to develop a multiplex PCR that could detect seven CPE genes simultaneously. The multiplex PCR was composed of seven primer sets for the detection of KPC, IMP, VIM, NDM-1, GES, OXA-23, and OXA-48. We designed different PCR product sizes of at least 100 bp. We evaluated the performance of this new test using 69 CPE-positive clinical isolates. Also, we confirmed the specificity to rule out false-positive reactions by using 71 carbapenem-susceptible clinical strains. Results: A total of 69 CPE clinical isolates showed positive results and were correctly identified as KPC (N=14), IMP (N=13), OXA-23 (N=12), OXA-48 (N=11), VIM (N=9), GES (N=5), and NDM (N=5) by the multiplex PCR. All 71 carbapenem-susceptible clinical isolates, including Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa, showed negative results. Conclusion: This multiplex PCR can detect seven CPE genes at a time and will be useful in clinical laboratories.

Behavior Analysis of Concrete Structure under Blast Loading : (II) Blast Loading Response of Ultra High Strength Concrete and Reactive Powder Concrete Slabs (폭발하중을 받는 콘크리트 구조물의 실험적 거동분석 : (II) 초고강도 콘크리트 및 RPC 슬래브의 실험결과)

  • Yi, Na Hyun;Kim, Sung Bae;Kim, Jang-Ho Jay;Cho, Yun Gu
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.29 no.5A
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    • pp.565-575
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    • 2009
  • In recent years, there have been numerous explosion-related accidents due to military and terrorist activities. Such incidents caused not only damages to structures but also human casualties, especially in urban areas. To protect structures and save human lives against explosion accidents, better understanding of the explosion effect on structures is needed. In an explosion, the blast load is applied to concrete structures as an impulsive load of extremely short duration with very high pressure and heat. Generally, concrete is known to have a relatively high blast resistance compared to other construction materials. However, normal strength concrete structures require higher strength to improve their resistance against impact and blast loads. Therefore, a new material with high-energy absorption capacity and high resistance to damage is needed for blast resistance design. Recently, Ultra High Strength Concrete(UHSC) and Reactive Powder Concrete(RPC) have been actively developed to significantly improve concrete strength. UHSC and RPC, can improve concrete strength, reduce member size and weight, and improve workability. High strength concrete are used to improve earthquake resistance and increase height and bridge span. Also, UHSC and RPC, can be implemented for blast resistance design of infrastructure susceptible to terror or impact such as 9.11 terror attack. Therefore, in this study, the blast tests are performed to investigate the behavior of UHSC and RPC slabs under blast loading. Blast wave characteristics including incident and reflected pressures as well as maximum and residual displacements and strains in steel and concrete surface are measured. Also, blast damages and failure modes were recorded for each specimen. From these tests, UHSC and RPC have shown to better blast explosions resistance compare to normal strength concrete.