• Title/Summary/Keyword: PCR 동정

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Isolation, Identification, and Fermentation Characteristics of Bacillus sp. with High Protease Activity from Traditional Cheonggukjang (전통 청국장으로부터 protease 분비능이 우수한 Bacillus sp. 균주의 분리 동정 및 발효 특성)

  • Ahn, Yong-Sun;Kim, Yong-Suk;Shin, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.38 no.1
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    • pp.82-87
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    • 2006
  • Twenty one strains strongly producing protease were isolated from Korean traditional Cheonggukjang. Eight strains selected by sensory evaluation on Cheonggukjang prepared with isolated strains were identified with based on biochemical properties a and 16S rDNA sequencing. Identified strains were Bacillus subtilis MB4, and Bacillus amyloliquefaciens A1, A2, B1, MC1, SB2, SC1, and SD1. Protease activities, important strain selection factor, were higher in Cheongjukjang prepared with B. subtilis MB4 (179.6 Unit) and B. amyloliquefaciens SB2 (201.9 Unit) than commercial traditional Cheonggukjang (97.9 Unit). Sensory evaluation revealed Cheonggukjang prepared with B. subtilis MB4 had flavor very similar to commercial traditional Cheonggukjang. Cheonggukjang prepared with B. suhtilis MB4 (0.0006 Pa s) and commercial traditional Cheonggukjang (0.0002 Pa s) revealed lower viscosities than those of Cheonggukjang prepared with B. amyloliquefaciens SB2, MC1, B1, A1, SD1, A2, and SC1 (0.006 to 0.008 Pa s at 1001/s. Results show Cheonggukjang could be prepared using single strain of B. subtilis MB4, maintaining high protease activity and very similar sensory and viscosity qualities with those of commercial traditional Cheonggukjang.

Molecular Genetic Characteristics of Methicillin-Resistant Staphylococcus aureus (MRSA) Isolated From Nasal Cavity of University Students (대학생들의 비강으로부터 분리된 메티실린 내성 황색포도알균의 분자유전학적 특성)

  • Lee Eun gwang;Oh Dae Hwan;Sunjin Jung;Sohyun Park;Yeonim Choi
    • Journal of the Health Care and Life Science
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    • v.9 no.2
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    • pp.303-308
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    • 2021
  • Staphylococcus aureus and methicillin resistant S. aureus (MRSA) is a major cause of nosocomial infections and is one of the most commonly isolated bacterial species in the hospital and continues to be an important pathogens in both community and hospital-acquires infection. The purpose of this study is to investigate the carrier rate of S. aureus and MRSA in the community and molecular genetic characteristics of these organisms. The identification of S. aureus and MRSA were done by the procedures in Murray's manual of Clinical Microbiology and antibiotic susceptibility patterns by the Clinical and Laboratory Standards Institute(CLSI). MRSA strains were confirms by oxacillin disk diffusion method. forty-six strains (71.9%) of S. aureus were isolated from the nasal specimens of 64 students in health science university. twenty-two strains (22%) of 46 S. aureus were resistant to penicillin and oxacillin. twenty-two strains of the 46 S. aureus isolates were methicillin-resistant Staphylococcus aureus (MRSA). The mecA genes in MRSA were detected by polymerase chain reaction (PCR). Community and nosocomial infections caused by methicillin-resistant Staphylococcus aureus are a significant problem worldwide. There continuous epidemiological study is to investigate the prevalence of MRSA in community acquired infections.

A Study on the Yogurt Manufacture Suitability and Antimicrobial Activity of Lactobacillus plantarum LHB55 Isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum LHB55의 항균성과 요구르트 제조 적합성 연구)

  • Lee, Seung-Gyu;Lee, Yeon-Jung;Kim, Min-Kyung;Han, Ki-Sung;Jeong, Seok-Geun;Oh, Mi-Hwa;Jang, Ae-Ra;Kim, Dong-Hun;Bae, In-Hyu;Ham, Jun-Sang
    • Journal of Animal Science and Technology
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    • v.52 no.2
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    • pp.141-148
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    • 2010
  • The aim of this study was to develop a new starter for fermented milk. The approach started with 103 acid-producing isolates from Kimchi, a type of spiced, fermented cabbage and then PCR screening was used to identify 72 Lactobacillus strains. The ability to inhibit the growth of food-borne human pathogens (Escherichia coli, Salmonella enteritidis, Staphylococcus aureus) of these strains were measured, using the paper disk method. Among them, one bacterium (LHB55) that showed a strong antibacterial activity against food-borne human pathogens was identified and further characterized, using 16S rDNA sequencing and API 50CHL system. Because this isolate was identified as L. plantarum, it was named as L. plantarum LHB55. The yogurt produced using commercial LAB with L. plantarum LHB55 did not display properties that are microbially or physico-chemically different from the control group, which suggests that L. plantarum LHB55 can be used as a useful starter for yogurt containing high antibacterial activity. We think that identifying effective starter strains enabling further development of fermented milk that can deliver better health benefits such as antimicrobial properties is of high significance, and thus our effort in this type of approach will continue.

Molecular Cloning and Expression Analysis of Red-spotted Grouper, Epinephelus akaara Hsp70 (수온변화에 따른 붉바리(Epinephelus akaara)의 heat shock protein (Hsp) 70 mRNA 발현)

  • Min, Byung Hwa;Hur, Jun Wook;Park, Hyung Jun
    • Journal of Life Science
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    • v.28 no.6
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    • pp.639-647
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    • 2018
  • A new heat shock protein 70 was identified in red-spotted grouper (Epinephelus akaara) based on an expression analysis. The cDNA of red-spotted grouper Hsp70 (designated RgHsp70) was cloned by the rapid amplification of cDNA ends (RACE) techniques. The full-length of RgHsp70 cDNA was 2,152 bp, consisting of a 5'-terminal untranslated region (UTR) of 105 bp, a 3'-terminal UTR of 274 bp, and an open reading frame (ORF) of 1,773 bp that encode a polypeptide of 590 amino acids with a theoretical molecular weight of 64.9 kDa and an estimated isoelectric point of 5.2. Multiple alignment and phylogenetic analyses revealed that the RgHsp70 gene shares a high similarity with other Hsp70 fish genes. RgHsp70 contained all three classical Hsp70 family signatures. The results indicated the RgHsp70 is a member of the heat shock protein 70 family. RgHsp70 mRNA was predominately expressed in the liver, with reduced expression noted in the head-kidney tissues. The expression analysis of different water temperatures (21, 18, 15 and $12^{\circ}C$) for sampled livers revealed that expression gradually increased at $12^{\circ}C$ compared to $21^{\circ}C$. In this study, the effects of water temperature lowering on the physiological conditions were investigated, and the results revealed that novel RgHsp70 may be an important molecule involved in stress responses.

Prevalence and Molecular Characterization of Methicillin Resistant Staphylococcus aureus Isolated from Raw Milk Samples in Gyeonggi-do, Korea (원유시료 중 메티실린 내성 황색포도알균의 분포 및 내성 유전자 특성 분석)

  • Kang, SoWon;Song, YoungCheon;Choi, SungSook
    • Journal of Food Hygiene and Safety
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    • v.29 no.3
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    • pp.223-227
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    • 2014
  • This study was investigated to determine the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) isolated from raw milk samples and to further study on the molecular characteristics of the MRSA isolates. Using Staphylococcus Medium 110, Staphylococcus spp. were isolated from raw milk samples and further identification was carried by Vitek2 system. Minimum inhibitory concentrations (MICs) of antibiotics were conducted by serial dilution method according to the Clinical Laboratory Standards Institute (CLSI) guideline. For the detection of resistance genes and molecular characterization, PCR reaction was performed by gene specific primers and followed by DNA sequencing. Of the 698 milk samples, 94 Staphylococcus aureus (S. aureus) were identified (94 S. aureus/286 Staphylococcus spp.). Of the 94 S. aureus, seven isolates have mecA, a methicillin resistant gene. mecA positive seven isolates were then characterized by staphylococcal cassette chromosome mec (SCCmec) typing, and Panton-Valentine Leukocidin (pvl) gene using PCR. All of mecA positive isolates were resistant to ampicillin and oxacillin, but sensitive to teicoplanin, vancomycin and ciprofloxacin. One of seven isolates was SCCmec type II and six isolates were type IV and all seven isolates were pvl gene negative.

Natural Occurrence of Fusarium Mycotoxins in Field-collected Maize and Rice in Korea in 2009 (2009년산 옥수수와 벼에서의 Fusarium 곰팡이독소 자연발생량 조사)

  • Lee, Seung-Ho;Son, Seung-Wan;Nam, Young-Ju;Shin, Jean-Young;Lee, Soo-Hyung;Kim, Mi-Ja;Yun, Jong-Chul;Ryu, Jae-Gee;Lee, Theresa
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.306-311
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    • 2010
  • To detect Fusarium mycotoxins, grain samples were collected from 32 rice fields all over the country and from 19 maize fields in eastern and midland provinces in Korea in 2009. Maize contamination with Fusarium species (54.9%) was higher than in rice (8.2%). Using Fusarium species specific PCR primer sets (Fg16 and VERT), 58 and 354 of total 506 isolates from maize samples were putatively identified as F. graminearum (11.5%) and F. verticillioides (70.0%), respectively. From rice samples, 276 of 315 isolates (87.8%) were putatively identified as F. graminearum but F. verticillioides was not identified. LC or LC-MS analysis of the samples revealed that fumonisin was the most commonly detected mycotoxin in maize samples but its level was below the regulation limit. Only two maize samples were contaminated with deoxynivalenol and zearalenone at the levels above the regulation limit. In rice samples, contamination with zearalenone was common but the levels were below the regulation limit. This study showed that most of the Korean maize and rice samples collected in 2009 were contaminated with Fusarium mycotoxins but the levels were below the Korean regulations for deoxynivalenol, fumonisin and zearalenone.

Some Characteristics of Melon necrotic spot virus-Me and Resistance Screen to the Virus in Melon Cultivars (멜론괴저반점바이러스-Me의 몇 가지 특성과 멜론 품종의 저항성 선발)

  • Choi, Gug-Seoun;Cho, Jeom-Deog;Chung, Bong-Nam;Cho, In-Sook;Kwon, Soon-Bae
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.254-258
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    • 2010
  • Melon necrotic spot virus (MNSV) is a very destructive disease to melon (Cucumis melo) plants. A MNSV was isolated from melon leaf showing necrotic spot symptoms at the plastic house in Naju, Korea in 2009. The isolate, designated as MNSV-Me, was identified and characterized by biological responses on several host plants, immuno captured RT-PCR and partial nucleotide sequencings of the genome. To evaluate MNSV-Me resistance in melon, thirty-five melon cultivars were mechanically inoculated on the cotyledon of the seedlings with the virus. MNSV-Me produced necrotic spots on the inoculated leaves of the all melon cultivars tested. Twenty-five cultivars were susceptible to the virus and they showed systemic necrotic spots on the leaves and/or necrosis longer than 3 cm in length on the stems within about forty days after inoculation. Five cultivars gave moderate resistance, no symptoms on the upper leaves but necrosis on the stem shorter than 3 cm in length. In an evaluation of MNSV-Me resistance in melon cultivars, 'Elstitan', 'Elsluxery', 'Betalichihage', 'Betalichi' and 'Womderfulhagae 1st' were found to have resistance by showing only faint necrosis on their stems.

A Survey of Viral Diseases of Proso Millet (Panicum miliaceum L.) and Sorghum (Sorghum bicolor L.) in South Korea (국내 기장 및 수수의 바이러스병 발생 조사)

  • Min, Hyun-Geun;Park, Chung Youl;Lee, Hong-Kyu;Yeom, Yoon-Ah;Oh, Jonghee;Kim, Bong-Sub;Lim, Seungmo;Yoon, Youngnam;Lee, Su-Heon
    • Research in Plant Disease
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    • v.23 no.3
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    • pp.262-267
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    • 2017
  • Throughout year 2015 to 2016, 101 proso millet and 200 sorghum samples were collected from five provinces in South Korea. The samples were subjected to paired-end RNA sequencing and further analyzed by RT-PCR. The results indicated that Rice black-streaked dwarf virus (RBSDV) was detected from sorghum collected in Gyeongsang province. The other four viruses, including RBSDV, Rice stripe virus (RSV), Barley virus G (BVG), and Cereal yellow dwarf virus (CYDV), were detected from proso millet. Among four viruses, both RSV and RBSDV were identified high frequency from proso millet collected from Gyeongsang province. Otherwise, BVG was nearly equally identified from five provinces, suggesting that the virus was supposedly widespread nationwide. RBSDV was first identified from both proso millet and sorghum in South Korea. The other virus annotated CYDV identified proso millet was shown to have relatively low identities compared to CYDV previously reported, suggesting that the virus might be new member of Polerovirus.

Distribution of Human Rotavirus Genotypes in a Tertiary Hospital, Seoul, Korea During 2009-2013 (2009년부터 2013년까지 서울의 일개 대학병원에서 동정된 로타바이러스 유전형의 분포)

  • Han, Tae Hee;Park, Sang-Hun;Chung, Ju-Young;Hwang, Eung-Soo
    • Pediatric Infection and Vaccine
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    • v.22 no.2
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    • pp.81-90
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    • 2015
  • Purpose: Group A rotavirus (RV) is most common etiologic agent of acute gastroenteritis (AGE) in children worldwide. Recently, vaccination has been introduced in several countries to reduce the disease burden caused by RV infections, but continuous surveillance of RV strains is necessary to detect the emergence of potential variants induced by vaccine-immune pressure. This study aimed to investigate the changing pattern of RV genotypes in children with AGE, following the introduction of vaccination in Korea. Methods: Genotyping of RVs by RT-PCR on the basis of VP7 and VP4 gene segment sequence was carried out on 201 rotavirus-positive stool samples, from children hospitalized with AGE between August 2009 and June 2013. We have directly sequenced PCR products and analyzed the phylogenetic tree. Results: The most prevalent G genotype was G9 (33.3%), followed by G1 (22.4%), G3 (15.9%), G2 (6.0%), G4 (3.0%), G10 (1.5%), and mixed G-type (15.4%), with some nontypeable cases (2.5%). The detected P genotypes were P[4] (45.3%), P[8] (43.8%), mixed P-type (10.4 %), and P[2] (0.5%). The G9P[4] genotype was predominantly observed in hospitalized cases in Seoul in 2010/2011, however G1P[8] has been re-emerged as the predominant genotype in the following season (P =0.004). Conclusions: It seems that the periodic fluctuation in predominance of the G1, G3, and G9 strains occurred in Korea during 2009-2013, following the introduction of RV vaccination.

Site-Specific Recombination by the Integrase MJ1 on Mammalian Cell (동물 세포 내에서 MJ1 인티그라제에 의한 부위 특이적 재조합)

  • Kim, Hye-Young;Yoon, Bo-Hyun;Chang, Hyo-Ihl
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.337-344
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    • 2011
  • Integrase MJ1 from the bacteriophage ${\Phi}FC1$ carries out recombination between two DNA sequences (the phage attachment site, attP and the bacterial attachment site, attB) in NIH3T3 mouse cells. In this study, the integration vector containing attP, attB and the integrase gene MJ, was constructed. The integration mediated by integrase MJ1 in Escherichia coli led to excision of LacZ. Therefore, the frequency of integration was measured by the counting of the white colony, which is detectable on X-Gal plates. The extrachromosomal integration in NIH3T3 mouse cells was monitored by the expression of the green fluorescent protein (GFP) as a reporter. To demonstrate integration mediated integrase MJ1 in NIH3T3 cells, vectors containing attP and attB were co-transfected into NIH3T3 cells. The integration was confirmed by fluorescent microscopy. The expression of GFP was induced in NIH3T3 cells expressing MJ1 without accessory factors. By contrast, the excision mediated by the MJ1 between attR and attL had no effect on the expression of GFP. These results suggest that integrase MJ1 may enable a variety of genomic modifications for research and therapeutic purposes in higher living cells.