• Title/Summary/Keyword: DNA염기

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Identification, sequence characterization and expression analysis of the arginine kinase gene in response to laminarin challenge from the Oriental land snail, Nesiohelix samarangae (동양달팽이(Nesiohelix samarangae)의 arginine kinase 유전자 분석 및 발현 패턴에 관한 연구)

  • Jeong, Ji Eun;Lee, Yong Seok
    • The Korean Journal of Malacology
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    • v.29 no.3
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    • pp.171-179
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    • 2013
  • Arginine kinase (ArK) is known to play an important role in most invertebrates the level of ATP by phosphorylation of phosphagens in cell and immuninty in living organisms. ArK has been identified in many kinds of organisms ranging from invertebrate to vertebrate. However, no ArK gene has been cloned and investigated from N. samarangae. This leads us to identify ArK cDNA (NsArK) from the expressed sequence tag (EST) sequencing of N. samarangae. Sequence analysis indicated that the coding region of 1,065 bp contains 355 amino acid residues. Molecular phylogenetic analysis shows that NsArK had very high similarities with mollusca and arthropoda. In an attempt to investigate a potential role of NsArK in the digestive gland of N. samarangae, expression patterns were analyzed. RT-PCR analsysis shows that NsArK mRNA is induced in the rane of 1.2 fold at 6 hr by laminarin when compared with the control. The immunnologial and physiological role of NsArK remains to be further investigated in N. samarangae.

Development of SCAR markers in Creeping bentgrass(Agrostis palustrics Huds.) cultivars (Creeping bentgrass(Agrostis palustrics Huds.) 품종별 SCAR markers 개발)

  • Jang, Duk-Hwan;Jung, Seung-Ho
    • Asian Journal of Turfgrass Science
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    • v.23 no.2
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    • pp.307-316
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    • 2009
  • Creeping bentgrass (Agrostis palustrics Huds.) is cool season turfgrasse that is used for putting green in golf course. Creeping bentgrass cultivars are difficult to distinguish with the same species because of similar morphological characters and low level of genetic diversity. The SCAR markers using the specific DNA can be useful for differentiating between creeping bentgrass cultivars. Five RAPD primers were used for specific band detection among creeping bentgrass cultivars, penncross, penn A-4, crenshaw, L-93, CY-2, T-1. The pairs of SCAR primers for six cultivers were designed by the specific sequences of the bands that amplified by RAPD. Three of the six SCAR primers could not make the use as SCAR primers because the specific false bands were detected in all cultivars. The remaining pairs of SCAR primer, CY850F/R, T700F/R, L2900F/R, amplified the specific band at expected size for three cultivars, CY-2, T-1, L-93, respectively. The CY850F/R primer amplified a band of 850bp in CY-2 cultivar, the T700F/R primer amplified a band of 700bp in T-1 cultivar, and the L2900F/R primer amplified a band of 2.9kb in L-93 cultivar. In this study we developed the SCAR markers to identify and distinguish the inerseeded creeping bentgrass cultivars in a golf course green.

Isolation of Egg-Contaminating Bacteria and Evaluation of Bacterial Radiation Sensitivity (계란 오염 세균의 분리 및 분리 균주의 감마선 감수성 평가)

  • Kim, Dong-Ho;Yun, Hye-Jeong;Song, Hyun-Pa;Lim, Byung-Lak;Jo, Cheo-Run
    • Food Science and Preservation
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    • v.15 no.5
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    • pp.774-781
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    • 2008
  • was performed and Staphylococcus sciuri, Bacillus cereus, Escherichia coli, Proteus mirabilis, and Enterococcus faecalis were identified. No Salmonella strain, a typical contaminant of eggs, was found. The radiation sensitivities of isolated bacteria and Salmonella typhimurium, in an inoculated model system, were expressed in $D_{10}$ values. The ranges of $D_{10}$ values shown by S. typhimurium, S. sciuri, B. cereus, E. coli, P. mirabilis, and E. faecalis were 0.365-0.399 kGy, 0.418-0.471 kGy, 1.075-1.119 kGy, 0.280-0.304 kGy, 1.132-1.330 kGy, and 0.993-1.290 kGy, respectively. The growth of all six test bacteria in eggs (inoculated at $10^6-10^7\;CFU/g$) during 3 days of post-irradiation storage at ambient conditions ($25^{\circ}C$) was recorded. S. typhimurium was eliminated by irradiation at 3 kGy, and E. coli and S. sciuri were eliminated by irradiation at 5 kGy. The viable cell counts of B. cereus, P. mirabilis, and E. faecalis in eggs showed 4-6 log reductions after irradiation with 5 kGy.

Analysis of Differentially Expressed Genes in Kiwifruit Actinidia chinensis var. 'Hongyang' (참다래 '홍양' 품종의 차등발현유전자 분석)

  • Bae, Kyung-Mi;Kwack, Yong-Bum;Shin, II-Sheob;Kim, Se-Hee;Kim, Jeong-Hee;Cho, Kang-Hee
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.448-456
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    • 2011
  • We used suppression subtractive hybridization (SSH) combined with mirror orientation selection (MOS) method to screen differentially expressed genes from red-fleshed kiwifruit 'Hongyang'. As a result, the 288 clones were obtained by subcloning PCR product and 192 clones that showed positive clones on colony PCR analysis were selected. All the positive clones were sequenced. After comparisons with the NCBI/Genbank database using the BLAST search revealed that 30 clones showed sequence similarity to genes from other organisms; 10 clones showed significant sequence similarity to known genes. Among these clones, 3 clones (AcF21, AcF42 and AcF106) had sequence homology to 1-aminicyclopropane-carboxylic acid (ACC)-oxidase (ACO) that known to be related to fruit ripening. The expression patterns of differentially expressed genes were further investigated to validate the SSH data by reverse transcription PCR (RT-PCR) and quantitative real-time PCR (qReal-time PCR) analysis. All the data from qReal-time PCR analysis coincide with the results obtained from RT-PCR analysis. Three clones were expressed at higher levels in 'Hongyang' than 'Hayward'. AcF21 was highly expressed in the other genes at 120 days after full bloom (DAFB) and 160 DAFB of 'Hongyang'.

Analysis of Genotype and Flanking Sequence Tagged from pooled Ds Insertional lines in rice (벼 Ds 삽입변이 pooling 계통들의 FST 및 유전자형 분석)

  • Ahn, Byung-Ohg;Kim, Jeong-Ho;Ji, Sang-hye;Yun, Doh-Won;Park, Yong-Hwan;Ji, Hyeon-So;Eun, Moo-Young;Lee, Gi-hwan;Suh, Seok-Cheol;Lee, Myung-Chul
    • Korean Journal of Breeding Science
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    • v.40 no.4
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    • pp.387-393
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    • 2008
  • Over 7 individual rice (Oryza sativa L.) plants per a line were sowed and sampled by pooled sampling method for genomic DNA extraction. The 5,400 flanking sequence tags (FSTs) were analysed by adaptor PCR and direct sequencing. FST analysis showed that the intragenic FSTs, the intergenic FSTs, and the original insertional sequences including hot spot covered 48.1% (2,597), 25.6% (1,383), and 25% (1,350), respectively. The 2,597 intragenic FSTs were used for genotyping to determine whether they are heterozygous or homozygous, and 1,393 core lines were selected. Among them, 422 knockout genes were distributed on chromosome 3, while 56 - 157 intragenic FSTs scattered on other chromosomes. Among 1,393 FSTs, known genes such as transcription factor covered 59.4% (827), while unknown genes such as expressed protein covered 40.6% (566). RT-PCR indicated that some core lines had no expression or decreased expression level in their knockout genes. It means that core lines are very useful knockout lines for functional genomic studies.

Protease Activity of Lactic Acid Bacteria Isolated from Korean Traditional Fermented Food (전통 발효식품으로부터 Protease 활성을 보유한 유산균의 분리 및 동정)

  • Kook, Moo Chang;Cho, Seok Cheol;Park, Hoon;Kim, Seung Seop;Pyun, Yu Ryang;Choi, Woon Yong;Lee, Hyeon Yong
    • Food Engineering Progress
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    • v.15 no.2
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    • pp.182-187
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    • 2011
  • A proteolytic lactic acid bacterium was isolated from Korean traditional fermented foods. The isolate BV-26, which had a protease activity (24 U/mg-crude protein), was identified as Lactobacillus plantarum by the API 50CHL kit and 16S rDNA analysis (99.9% of homology), and named as L. plantarum BV-26. Cell growth and protease activity of L. plantarum BV-26 was determined in MRS broth using 5L jar fermentor at $30^{\circ}C$. The maximum growth of L. plantarum BV-26 was reached at 18 hr in MRS broth, while protease activity of BV-26 was detectable at 12 hr and the highest activity was obtained after 16 hr cultivation. Therefore, we expect that the proteolytic lactic acid bacteria, L. plantarum BV-26, may be used as a starter for the fermentation of animal feed. Especially, the fermentation of soybean meal with the strain can be applied for improving feed utilization.

Breeding of F1 Hybrid for Oriental Mustard(Brassica juncea L. Czern) Using the Cytoplasmic Male Sterile Line (웅성불임 인자를 이용한 갓(Brassica juncea L. Czern)의 F1 육종)

  • Park, Yong Ju;Min, Byung Whan
    • Journal of agriculture & life science
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    • v.52 no.6
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    • pp.27-36
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    • 2018
  • Recently, the demand for new cultivar of oriental mustard(Brassica juncea) is increased as the consumption of oriental mustard has increased dramatically in the market due to Kimchi is attracting the world's attention. However, absence of seed supply system can cause many problems including inbreeding depression due to self seed production, deterioration of the seed purity and heterogeneity of commercial seed. To establish the $F_1$ variety breeding system of oriental mustard, pure line and inbred lines were screened from inbreeding genetic resources. Male-sterile line was also selected from breeding combinations for the quality improvement of mustard. The combining ability from(Indojasai ${\times}$ Goheungdamyang) combinations and isolation line of(MS910 ${\times}$ Japan red mustard 8 ${\times}$ Ganghwa mustard 9) was highest, thus these lines were selected as parental lines. PCR(Polymerase Chain Reaction) and gene sequence analysis revealed that the genes related to CMS(orf263, orf220, and orf288) were distributed in mitochondria. The isolated lines from this study also showed good performance in yield test and farmhouse prove test.

Molecular Analysis of Carbapenem-Resistant Pseudomonas aeruginosa Isolated from Patients Hospitalized in Daejeon between 2008 and 2014 Years (대전지역의 입원환자에서 분리된 Carbapenem 내성 Pseudomonas aeruginosa의 분자역학조사(2008년에서 2014년까지))

  • Cho, Hye Hyun
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.4
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    • pp.406-413
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    • 2018
  • The emergence of carbapenem resistance among Pseudomonas aeruginosa has become an increasing problem worldwide. In particular, $metallo-{\beta}-lactamases$ (MBLs) are responsible for the high-level resistance to carbapenem. Sequence type 235 (ST235) has been found internationally in a multidrug-resistant clone and is involved in the dissemination of genes encoding IMP-6 and VIM-2. This study examined the prevalence of MBLs and the epidemiological relationship in carbapenem-resistant P. aeruginosa (CRPA) isolates obtained from a tertiary hospital in Daejeon, Korea, between March 2008 and June 2014. The antimicrobial susceptibilities were determined using the disk-diffusion method and PCR and DNA sequencing were used to identify the MBL genes. In addition, an epidemiological relationship was investigated by multilocus sequence typing (MLST). Among the 110 CRPA isolates, 32 isolates (29.1%) were MBL-producers; the major type was IMP-6 (29 isolates, 90.6%). VIM-2 was identified in 3 isolates (9.4%) of ST357. IMP-6-producing isolates were multidrug-resistant (MDR) and belonged to ST235. ST235 (55 isolates, 50.0%) was the clone most frequently detected and has gradually emerged during a seven-year period. To prevent the spread of MDR ST235 P. aeruginosa isolates, the current widespread use of carbapenems needs to be curtailed, and novel continuous monitoring strategies should be developed as soon as possible.

Isolation and Identification of Ampicillin-resistant Bacteria in Changwon (창원근교에서의 ampicillin 내성세균의 분리 및 동정)

  • Bae, Young-Min
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1529-1535
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    • 2018
  • The number of antibiotic-resistant bacteria is increasing rapidly while the discovery rate of new antibiotics is in decline. A systematic study is therefore necessary to investigate which bacteria are resistant to medically important antibiotics and how high that resistance is. To that end, this study aimed to analyze which bacteria demonstrated resistance to ampicillin, one of the currently most-widely used medical antibiotics. Water samples were collected from the Changwon-Cheon that runs through Changwon City and from the pond in front of the dormitory building at Changwon University. Hundreds of ampicillin-resistant colonies were obtained and 22 morphologically distinct examples were chosen for further study. These bacteria were identified by amplifying their 16S rRNA genes and comparing those sequences with data in GenBank. The bacteria was identified as belonging to 10 families, 12 genera, and 17 species, and all were able to grow in the presence of $50{\mu}g/ml$ ampicillin while seven showed growth at ampicillin concentrations as high as 1.5 mg/ml.

Molecular Detection of Virulence Factors in Carbapenem-Resistant Pseudomonas aeruginosa Isolated from a Tertiary Hospital in Daejeon (대전지역의 3차 병원에서 분리된 Carbapenem 내성 Pseudomonas aeruginosa의 병독성 인자 검출)

  • Cho, Hye Hyun
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.301-308
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    • 2019
  • The emergence and spread of multidrug resistant (MDR) Pseudomonas aeruginosa is a critical problem worldwide. The pathogenesis of P. aeruginosa is due partly to the production of several cell-associated and extracellular virulence factors. This study examined the distribution of virulence factors and antimicrobial resistance patterns of carbapenem-resistant P. aeruginosa (CRPA) isolated from a tertiary hospital in Daejeon, Korea. Antimicrobial susceptibility testing was performed using the disk diffusion method, and PCR and DNA sequencing were performed to determine for the presence of virulence genes. In addition, the sequence type (ST) of MDR P. aeruginosa was investigated by multilocus sequence typing (MLST). Among 32 CRPA isolates, 14 (43.8%) were MDR and the major ST was ST235 (10 isolates, 71.4%). All isolates were positive for the presence of virulence genes and the most prevalent virulence genes were toxA, plcN, and phzM (100%). All isolates carried at least eight or more different virulence genes and nine (28.1%) isolates had 15 virulence genes. The presence of the exoU gene was detected in 71.4% of the MDR P. aeruginosa isolates. These results indicate that the presence of the exoU gene can be a predictive marker for the persistence of MDR P. aeruginosa isolates.