• 제목/요약/키워드: DNA storage

검색결과 186건 처리시간 0.024초

Comparative Whole Cell Proteomics of Listeria monocytogenes at Different Growth Temperatures

  • Won, Soyoon;Lee, Jeongmin;Kim, Jieun;Choi, Hyungseok;Kim, Jaehan
    • Journal of Microbiology and Biotechnology
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    • 제30권2호
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    • pp.259-270
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    • 2020
  • Listeria monocytogenes is a gram-positive, facultative anaerobe food pathogen responsible for the listeriosis that mostly occurs during the low-temperature storage of a cold cut or dairy products. To understand the systemic response to a wide range of growth temperatures, L. monocytogenes were cultivated at a different temperature from 10℃ to 42℃, then whole cell proteomic analysis has been performed both exponential and stationary cells. The specific growth rate increased proportionally with the increase in growth temperature. The maximum growth rate was observed at 37℃ and was maintained at 42℃. Global protein expression profiles mainly depended on the growth temperatures showing similar clusters between exponential and stationary phases. Expressed proteins were categorized by their belonging metabolic systems and then, evaluated the change of expression level in regard to the growth temperature and stages. DnaK, GroEL, GroES, GrpE, and CspB, which were the heat&cold shock response proteins, increased their expression with increasing the growth temperatures. In particular, GroES and CspB were expressed more than 100-fold than at low temperatures during the exponential phase. Meanwhile, CspL, another cold shock protein, overexpressed at a low temperature then exponentially decreased its expression to 65-folds. Chemotaxis protein CheV and flagella proteins were highly expressed at low temperatures and stationary phases. Housekeeping proteins maintained their expression levels constant regardless of growth temperature or growth phases. Most of the growth related proteins, which include central carbon catabolic enzymes, were highly expressed at 30℃ then decreased sharply at high growth temperatures.

Transgenic Tobacco Plants Expressing the Bacterial Levansucrase Gene Show Enhanced Tolerance to Osmotic Stress

  • Park, Jeong-Mee;Kwon, Suk-Yoon;Song, Ki-Bang;Kwak, Ju-Won;Lee, Suk-Bae;Nam, Young-Woo;Shin, Jeong-Sheop;Park, Young-In;Rhee, Sang-Ki;Paek, Kyung-Hee
    • Journal of Microbiology and Biotechnology
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    • 제9권2호
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    • pp.213-218
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    • 1999
  • Fructans are polyfructose molecules that function as nonstructural storage carbohydrates in several plants. In addition, it has been suggested that, due to their solubility, they can play an important role in helping plants survive periods of osmotic stress. In order to study the effect of levan synthesis on plant growth, the coding region of the levansucrase gene, which was isolated from Zymomonas mobilis, was introduced into tobacco plants using Agrobacterium tumefaciens-mediated transformation. The presence of the levansucrase gene in transgenic plants was verified by genomic DNA gel blot analysis. RNA gel blot and immunoblot analyses showed an accumulation of the corresponding transcript and protein product of the bacterial levansucrase gene in transgenic plants. Furthermore, a thin layer chromatography analysis revealed that fructans were synthesized and deposited in transgenic tobacco plants. When $T_1$ seeds were germinated and grown under polyethylene glycol-mediated drought stress or cold stress, the transgenic seedlings displayed a substantially higher level of growth than that of untransformed plants. These results suggest that fructans may playa significant role in the tolerance of plants under osmotic stress.

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Sulfate Reduction for Bioremediation of AMD Facilitated by an Indigenous Acid- and Metal-Tolerant Sulfate-Reducer

  • Nguyen, Hai Thi;Nguyen, Huong Lan;Nguyen, Minh Hong;Nguyen, Thao Kim Nu;Dinh, Hang Thuy
    • Journal of Microbiology and Biotechnology
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    • 제30권7호
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    • pp.1005-1012
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    • 2020
  • Acid mine drainage (AMD) has been a serious environmental issue that threatens soil and aquatic ecosystems. In this study, an acid-tolerant sulfate-reducing bacterium, strain S4, was isolated from the mud of an AMD storage pond in Vietnam via enrichment in anoxic mineral medium at pH 5. Comparative analyses of sequences of the 16S rRNA gene and dsrB gene involved in sulfate reduction revealed that the isolate belonged to the genus Desulfovibrio, and is most closely related to Desulfovibrio oxamicus (with 99% homology in 16S rDNA sequence and 98% homology in dsrB gene sequence). Denaturing gradient gel electrophoresis (DGGE) analyses of dsrB gene showed that strain S4 represented one of the two most abundant groups developed in the enrichment culture. Notably, strain S4 was capable of reducing sulfate in low pH environments (from 2 and above), and resistance to extremely high concentration of heavy metals (Fe 3,000 mg/l, Zn 100 mg/l, Cu 100 mg/l). In a batch incubation experiment in synthetic AMD with pH 3.5, strain S4 showed strong effects in facilitating growth of a neutrophilic, metal sensitive Desulfovibrio sp. strain SR4H, which was not capable of growing alone in such an environment. Thus, it is postulated that under extreme conditions such as an AMD environment, acid- and metal-tolerant sulfate-reducing bacteria (SRB)-like strain S4 would facilitate the growth of other widely distributed SRB by starting to reduce sulfate at low pH, thus increasing pH and lowering the metal concentration in the environment. Owing to such unique physiological characteristics, strain S4 shows great potential for application in sustainable remediation of AMD.

Growth Retardation and Death of Rice Plants Irradiated with Carbon Ion Beams Is Preceded by Very Early Dose- and Time-dependent Gene Expression Changes

  • Rakwal, Randeep;Kimura, Shinzo;Shibato, Junko;Nojima, Kumie;Kim, Yeon-Ki;Nahm, Baek Hie;Jwa, Nam-Soo;Endo, Satoru;Tanaka, Kenichi;Iwahashi, Hitoshi
    • Molecules and Cells
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    • 제25권2호
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    • pp.272-278
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    • 2008
  • The carbon-ion beam (CIB) generated by the heavy-ion medical accelerator in Chiba (HIMAC) was targeted to 7-day-old rice. Physiological parameters such as growth, and gene expression profiles were examined immediately after CIB irradiation. Dose-dependent growth suppression was seen three days post-irradiation (PI), and all the irradiated plants died by 15 days PI. Microarray (Agilent rice 22K) analysis of the plants immediately after irradiation (iai) revealed effects on gene expression at 270 Gy; 353 genes were up-regulated and 87 down-regulated. Exactly the same set of genes was affected at 90 Gy. Among the highly induced genes were genes involved in information storage and processing, cellular processes and signaling, and metabolism. RT-PCR analysis confirmed the microarray data.

Lycopene의 사료 내 첨가가 육계와 산란계의 항산화에 미치는 영향 (Effects of Dietary Lycopene Supplementation on Antioxidtion in Broiler and Layer)

  • 주원돈;안병기;강창원
    • 한국가금학회:학술대회논문집
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    • 한국가금학회 2005년도 제22차 정기총회 및 학술발표회
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    • pp.17-30
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    • 2005
  • Lycopene is the red-coloured carotenoid predominantly found in tomato fruit and one of the major carotenoids in the diets of North American and Europeans. Interest in lycopene is growing rapidly following the recent publication of its effects as a natural antioxidant and prevention of cardiovascular disease and cancers. Lycopene, a polyene hydrocarbon carotenoid haying 13 double bond, of which 11 are conjugated double bonds in a linear array exhibits a strong antioxidant property almost twice as strong as that of ${\beta}$-carotene. Lycopene has been shown in recent epidemiological and experimental studies to protect against oxidative damage of DNA which plays an important part in development of various cancer. Lycopene also contribute towards reducing the risk of cardiovascular diseases by preventing oxidation of low-density lipoprotein(LDL) cholesterol. This review summarize our knowledge and the current understanding of lycopene in human health as well as the results of experiments we conducted. We conducted experiments for investigating the effects of antioxidant in broiler and the possibilities of production of high quality eggs containing lycopene by the dietary lycopene supplementation with synthetic lycopene or tomato paste. The results shows that thiobarbituric acid reaction substances(TBARS) values in process of LDL oxidation in blood serum of broiler were significantly decreased by dietary lycopene and tomato paste. The dietary lycopene supplementation resulted in improved egg yolk color and in decreased the malondialdehyde (MDA) of egg yolk after 4 wk of storage at room temperature significantly(P<0.05). The dietary tomato paste was more effective in the MDA reduction compared to the lycopene(P<0.05). The contents of lycopene in egg yolk of the lycopene supplementation groups were significantly higher than those of the control group.

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Development of Anti-Obesity Agent from Resource Plants

  • Jeong, Yong-Joon;Kang, Se-Chan
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2012년도 정기총회 및 춘계학술발표회
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    • pp.15-15
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    • 2012
  • Obesity is a physical condition that results from excessive storage of fat in the body. The present study examined the anti-obesity effects of the selected natural medicine, Galla rhois extract (GRE) and solvent fractions on 3T3-L1 preadipocytes and in vivo studies. Here, we show that EtOAc fraction of Galla rhois inhibits the differentiation of the 3T3-L1 preadipocytes induced by differentiated medium in a dose-dependent manner. To investigate the effect of the GRE-EtOAc fraction on obesity in high fat diet-fed C57BL/6 mice, which included a normal diet (ND), high-fat diet (HFD) and HFD+GRE concentration-dependent, were fed to the mice for 6 weeks. The GRE-EtOAc fraction was inhibited the highest adipocyte differentiation in vitro, the GRE supplement significantly decreased body weight and visceral fat mass compared to the HFD group. The total cholesterol and triglyceride levels in the plasma were significantly decreased by GRE supplementation compared with those of the HFD group. Also, we aimed to determine the differentiation inhibition and the modulation of differentiation genes brought about by the Galla rhois in adipocyte. A cDNA microarray-based method was introduced for the high contents screening (HCS) of gene expressions. This technology has revolutionized gene expression studies by providing the means to measure mRNA levels in thousands of genes simultaneously in simple and complex biological samples. 13 genes were founded to be affected in their expression levels by more than 5-fold up-regulation after 4 days treatment with the EtOAc fraction from Galla rhois. Otherwise, 21 genes were founded to be affected in their expression levels by more than 5-fold down-regulation treated with the EtOAc fraction. Therefore, Galla rhois extract may be considered for use in a therapeutic agent to control obesity.

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Label-free Femtomolar Detection of Cancer Biomarker by Reduced Graphene Oxide Field-effect Transistor

  • Kim, Duck-Jin;Sohn, Il-Yung;Jung, Jin-Heak;Yoon, Ok-Ja;Lee, N.E.;Park, Joon-Shik
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2012년도 제42회 동계 정기 학술대회 초록집
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    • pp.549-549
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    • 2012
  • Early detection of cancer biomarkers in the blood is of vital importance for reducing the mortality and morbidity in a number of cancers. From this point of view, immunosensors based on nanowire (NW) and carbon nanotube (CNT) field-effect transistors (FETs) that allow the ultra-sensitive, highly specific, and label-free electrical detection of biomarkers received much attention. Nevertheless 1D nano-FET biosensors showed high performance, several challenges remain to be resolved for the uncomplicated, reproducible, low-cost and high-throughput nanofabrication. Recently, two-dimensional (2D) graphene and reduced GO (RGO) nanosheets or films find widespread applications such as clean energy storage and conversion devices, optical detector, field-effect transistors, electromechanical resonators, and chemical & biological sensors. In particular, the graphene- and RGO-FETs devices are very promising for sensing applications because of advantages including large detection area, low noise level in solution, ease of fabrication, and the high sensitivity to ions and biomolecules comparable to 1D nano-FETs. Even though a limited number of biosensor applications including chemical vapor deposition (CVD) grown graphene film for DNA detection, single-layer graphene for protein detection and single-layer graphene or solution-processed RGO film for cell monitoring have been reported, development of facile fabrication methods and full understanding of sensing mechanism are still lacking. Furthermore, there have been no reports on demonstration of ultrasensitive electrical detection of a cancer biomarker using the graphene- or RGO-FET. Here we describe scalable and facile fabrication of reduced graphene oxide FET (RGO-FET) with the capability of label-free, ultrasensitive electrical detection of a cancer biomarker, prostate specific antigen/${\alpha}$ 1-antichymotrypsin (PSA-ACT) complex, in which the ultrathin RGO channel was formed by a uniform self-assembly of two-dimensional RGO nanosheets, and also we will discuss about the immunosensing mechanism.

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마이크로어레이 데이터의 구조적 유사성을 이용한 효율적인 저장 구조의 설계 (Design of Efficient Storage Exploiting Structural Similarity in Microarray Data)

  • 윤종한;신동규;신동일
    • 정보처리학회논문지D
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    • 제16D권5호
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    • pp.643-650
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    • 2009
  • 생명정보 대량 획득기술의 하나인 마이크로어레이(microarray)는 DNA와 각종 유전자 연구에 사용되는 도구로 확립되면서, 생명정보학(Bioinformatics)분야의 발전에 크게 기여하였다. 그러나 마이크로어레이는 생명정보학분야의 핵심기술 중 하나로 발전하였음에도 불구하고 실험으로 생성되는 데이터는 형태가 다양하고 매우 복잡한 형태를 갖기 때문에 데이터의 공유나 저장에서 많은 어려움을 겪고 있다. 본 논문에서는 마이크로어레이 데이터의 관리를 원활하게 하기위한 XML 기반의 표준 포맷인 MAGE-ML스키마에서 구조적으로 유사한 엘리먼트가 반복적으로 나타나는 특징과 대다수의 엘리먼트들이 특정 엘리먼트의 자식으로만 온다는 구조적 특징을 이용하여, MAGE-ML의 스키마를 단순화 하고 저장구조를 효율적으로 설계하는 방법을 제안한다. 이 방법에서 인라인 기법(Inlining Technique)을 이용한 스키마의 단순화와 새롭게 제시하는 엘리먼트의 구조적 형태를 기준으로 분류하는 기법을 이용한다. 이를 통하여 데이터베이스 스키마는 간략화 되며 테이블조인의 횟수가 줄어들고 성능은 향상된다.

The Role of Rice Vacuolar Invertase2 in Seed Size Control

  • Lee, Dae-Woo;Lee, Sang-Kyu;Rahman, Md Mizanor;Kim, Yu-Jin;Zhang, Dabing;Jeon, Jong-Seong
    • Molecules and Cells
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    • 제42권10호
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    • pp.711-720
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    • 2019
  • Sink strength optimizes sucrose import, which is fundamental to support developing seed grains and increase crop yields, including those of rice (Oryza sativa). In this regard, little is known about the function of vacuolar invertase (VIN) in controlling sink strength and thereby seed size. Here, in rice we analyzed mutants of two VINs, OsVIN1 and OsVIN2, to examine their role during seed development. In a phenotypic analysis of the T-DNA insertion mutants, only the OsVIN2 mutant osvin2-1 exhibited reduced seed size and grain weight. Scanning electron microscopy analysis revealed that the small seed grains of osvin2-1 can be attributed to a reduction in spikelet size. A significant decrease in VIN activity and hexose level in the osvin2-1 spikelets interfered with spikelet growth. In addition, significant reduction in starch and increase in sucrose, which are characteristic features of reduced turnover and flux of sucrose due to impaired sink strength, were evident in the pre-storage stage of osvin2-1 developing grains. In situ hybridization analysis found that expression of OsVIN2 was predominant in the endocarp of developing grains. A genetically complemented line with a native genomic clone of OsVIN2 rescued reduced VIN activity and seed size. Two additional mutants, osvin2-2 and osvin2-3 generated by the CRISPR/Cas9 method, exhibited phenotypes similar to those of osvin2-1 in spikelet and seed size, VIN activity, and sugar metabolites. These results clearly demonstrate an important role of OsVIN2 as sink strength modulator that is critical for the maintenance of sucrose flux into developing seed grains.

Development of SSR Markers and Their Use in Studying Genetic Diversity and Population of Finger Millet (Eleusine coracana L. Gaertn.)

  • Lee, Kyung Jun;Yoon, Mun-Sup;Shin, Myoung-Jae;Lee, Jung-Ro;Cho, Yang-Hee;Lee, Ho-Sun;Ma, Kyung-Ho;Lee, Gi-An
    • Plant Breeding and Biotechnology
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    • 제5권3호
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    • pp.183-191
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    • 2017
  • Finger millet (Eleusine coracana L. Gaertn.) is an important cereal crop in eastern Africa and southern India with excellent grain storage capacity and the unique ability to thrive in extreme environmental conditions. In this study, we analyzed the genetic diversity and population structure of finger millet using 12 developed microsatellites. By sequencing 815 clones from an SSR-enriched genomic DNA library, we obtained 12 polymorphic SSR markers, which also revealed successful amplicons in finger millet accessions. Using the developed SSR markers, we estimated genetic diversity and population structure among 76 finger millet accessions in Asia, Africa, and unknown origins. The number of alleles ranged from 2 to 9, with an average of 3.3 alleles. The mean values of observed heterozygosity and expected heterozygosity were 0.27 and 0.35, respectively. The average polymorphism information content was 0.301 in all 76 finger millet accessions. AMOVA analysis showed that the percentage of molecular variance among the populations was 1%, that among individuals was 5%, and that within individuals was 94%. In STRUCTURE analysis, the 76 finger millet accessions were divided into two subpopulations which had an admixture of alleles. There was a correspondence among PCoA, AMOVA, and population structure. This study may form the basis for a finger millet breeding and improvement program.