• 제목/요약/키워드: molecular biological experiment

검색결과 93건 처리시간 0.033초

이온성 액체의 셀룰로오스 용해성 개선을 위한 전자빔 처리 효과 (Improvement in Dissolution of Cellulose with Ionic liquid by the Electron Beam Treatment)

  • 이원실;정웅기;성용주
    • 펄프종이기술
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    • 제45권2호
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    • pp.56-65
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    • 2013
  • Electron beam treatment was applied for improving dissolution of cellulose with ionic liquids. Two ionic liquids, 1-allyl-3-methylimidazolium chloride ([Amim]Cl]: AC) and 1,3-dimethylimidazolium methylphosphite ([Dmim][$(MeO)(H)PO_2$]: Me) were used for this experiment. Treatment with electron beams up to dose of 400 kGy resulted in the increase of hot water extract and alkali extract of cotton pulp and the great reduction in the molecular weight of cellulose. For the dissolution of cotton pulp with two ionic liquids, the electron beam treated samples showed faster dissolution. The dissolved cellulose with Me ionic liquid were regenerated with acetonitrile and the structure of regenerated cellulose showed distinct difference depending on the electron beam treatment. Those results provide the electron beam pre-treatment could be applied as an energy efficient and environmentally benign method to increase the dissolution of cellulose with ionic liquids.

초음파 및 침적방법을 이용한 강황 (Curcuma longa)으로부터 Curcuminoids의 확인 (Identification of Curcuminoids from Turmeric (Curcuma longa) Using Ultrasonic Wave and Dipping Method)

  • 이광진;마진열;김영식
    • KSBB Journal
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    • 제27권1호
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    • pp.33-39
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    • 2012
  • In this study three major curcuminoids in turmeric curcumin (1), demethoxycurcumin (2) and bismethoxycurcumin (3) were efficiently extracted by optimizing extraction condition and simultaneously identified using a fast and reliable RP-HPLC-UV-MS and TLC method. The analysis by the $C_{18}$ column was performed and the UV wavelength was fixed at 425 nm. In this result, the total extraction yield of turmeric (Curcuma longa) was increased with extraction time from 1 to 7 h. So, optimum extraction time is 4 h. Also, the highest yield of extraction amount 0.433g 8.66% was obtained by ultrasonic waves with quarter frequency kHz and an extraction time of 7 h. The experiment method was consistent with theoretical Value $r^2$ = 0.987 (1), 0.997 (2) and 0.998 (3). Moreover, LC-MS analysis provided efficiently molecular weight information of three major curcuminoids in turmeric extracts and high purity (~95%) of the curcuminoids were obtained. This work offers would be useful for chemical and biological studies of natural plants and its products.

Screening of Antagonistic Bacillus against Brown Rot in Dendrocalamus latiflorus and Preparation of Applying Bacterial Suspension

  • Fengying Luo;Hang Chen;Wenjian Wei;Han Liu;Youzhong Chen;Shujiang Li
    • The Plant Pathology Journal
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    • 제40권1호
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    • pp.1-15
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    • 2024
  • The aim of this study was to isolate biocontrol bacteria that could antagonize brown rot of Dendrocalamus latiflorus, optimize the culture conditions, and develop an effective biocontrol preparation for brown rot of D. latiflorus. This study isolated a bacterium with an antagonistic effect on bamboo brown rot from healthy D. latiflorus rhizosphere soil. Morphology, molecular biology, and physiological biochemistry methods identified it as Bacillus siamensis. The following culturing media and conditions improved the inhibition effect of B. siamensis: the best culturing media were 2% sucrose, 1.5% yeast extract, and 0.7% potassium chloride; the optimal culturing time, temperature, pH, and inoculation amount were 48 h, 30℃, 6, and 20%. The optimum formula of the applying bacterial suspension was 14% sodium dodecyl benzene sulfonate emulsifier, 4% Na2HPO4·2H2O, 0.3% hydroxypropyl methylcellulose thickener, and 20% B. siamensis. The pot experiment results showed the control effect of applying bacterial suspension, diluted 1,000 times is still better than that of 24% fenbuconazole suspension. The applying bacterial suspension enables reliable control of brown rot in D. latiflorus.

Biochemical Characteristics of an Alanine Racemase from Xanthomonas oryzae pv. oryzae

  • Kang, Han-Chul;Yoon, Sang-Hong;Lee, Chang-Muk;Koo, Bon-Sung
    • Journal of Applied Biological Chemistry
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    • 제54권4호
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    • pp.231-237
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    • 2011
  • A gene encoding a putative alanine racemase in Xanthomonas. oryzae pv. oryzae was cloned, expressed and characterized. Expression of the cloned gene was performed in Escherichia coli BL21(DE3)pLys using a pET-21(a) vector harbouring $6{\times}histidine$ tag. Purification of the recombinant alanine racemase by affinity chromatography resulted in major one band by sodium dodecyl sulfate polyacryl amide gel electrophoresis analysis, showing about 45 kDa of molecular weight. The alanine racemase gene, cloned in this experiment, appears to be constitutively expressed in X. oryzae, as analyzed by reverse transcriptase polymerase chain reaction. The enzyme was the most active toward L-alanine and secondly D-alanine, showing a racemic reaction, thus the enzyme is considered as an alanine racemase. The enzyme was considerably activated by addition of pyridoxal-5-phosphate (PLP), showing that 75% increase in activity was observed at 0.3 mM, compared with control. D-Cysteine as well as L-cysteine significantly inhibited the enzyme activity. The inhibitions by cysteines were more prominent in the absence of PLP, showing 9 and 5% of control activity at 2 mM of addition, respectively. The enzyme was the most active at pH 8.0 and more stable at alkaline pHs than acidic pH condition.

Panaxadiol saponins treatment caused the subtle variations in the global transcriptional state of Asiatic corn borer, Ostrinia furnacalis

  • Liu, Shuangli;Xu, Yonghua;Gao, Yugang;Zhao, Yan;Zhang, Aihua;Zang, Liansheng;Wu, Chunsheng;Zhang, Lianxue
    • Journal of Ginseng Research
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    • 제44권1호
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    • pp.123-134
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    • 2020
  • Background: The lepidopteran Asiatic corn borer (ACB), Ostrinia furnacalis (Guenee), has caused huge economic losses throughout the Asian-Western Pacific region. Usually, chemical pesticides are used for the control, but excessive use of pesticides has caused great harm. Therefore, the inartificial ecotypic pesticides to ACB are extremely essential. In our previous study, we found that panaxadiol saponins (PDS) can effectively reduce the harm of ACB by causing antifeedant activity. Therefore, it is necessary to reveal the biological molecular changes in ACB and the functionary mechanism of PDS. Methods: We analyzed the global transcription of ACB with different PDS concentration treatment (5 mg/mL, 10 mg/mL, and 25 mg/mL) by high-throughput sequencing and de novo transcriptome assembly method. Results: PDS treatment could cause the changes of many gene expressions which regulate its signal pathways. The genes in peroxisome proliferator-activated receptor (PPAR) signaling pathway were significantly downregulated, and then, the downstream fatty acid degradation pathway had also been greatly affected. Conclusion: Through this experiment, we hypothesized that the occurrence of antifeedant action of ACB is because the PDS brought about the downregulation of FATP and FABP, the key regulators in the PPAR, and the downregulation of FATP and FABP exerts further effects on the expression of SCD-1, ACBP, LPL, SCP-X, and ACO, which leads to the disorder of PPAR signaling pathway and the fatty acid degradation pathway. Not only that, PDS treatment leads to enzyme activity decrease by inhibiting the expression of genes associated with catalytic activity, such as cytochrome P450 and other similar genes.

마이크로어레이 데이터의 게놈수준 분석을 위한 퍼지 패턴 매칭에 의한 유전자 필터링 (Gene filtering based on fuzzy pattern matching for whole genome micro array data analysis)

  • 이선아;이건명;이승주;김원재;김용준;배석철
    • 한국지능시스템학회논문지
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    • 제18권4호
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    • pp.471-475
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    • 2008
  • 생명과학분야에서 마이크로어레이 기술은 세포에서의 RNA 발현 프로파일을 관찰할 수 있도록 함으로써 생명현상의 규명 및 약물개발 등에서 분자수준의 생명현상에 대한 관찰과 분석이 가능해지고 있다. 마이크로어레이 데이터분석에서는 특정한 처리나 과정에서 현저한 특성을 보이는 유전자를 식별하기 위한 분석뿐만 아니라 유전자 전체인 게놈수준에서의 분석도 이루어진다. 약물반응 실험에서는 약물에 대한 게놈수준의 발현 프로파일을 관찰하는 것도 많은 정보를 제공할 수 있다. 약물실험에서는 대조군과 실험군들간에 관심있는 상대적인 발현특성을 갖는 유전자군을 전체적으로 추출하는 것이 필요한 경우가 있다. 예를 들면 정상군은 두개의 실험군에 대해서 중간정도의 발현정도를 갖는 유전자군을 식별하는 분석을 하는 경우, 생물학적인 데이터의 특성상 절대값을 비교하는 방법으로는 유용한 유전자들을 효과적으로 식별해 낼 수 없다. 이 논문에서는 정상군과 실험군들의 발현정도값의 경향을 판단하기 위해서 각 유전자에 대해서 집단별 대표값을 선정하여 퍼지집합으로 집단의 값의 범위를 결정하고, 이를 이용하여 특성 패턴을 갖는 유전자들을 식별해내는 방법을 제안하고, 실제 데이터를 통해서 실험한 결과를 보인다.

CLUSTERING DNA MICROARRAY DATA BY STOCHASTIC ALGORITHM

  • Shon, Ho-Sun;Kim, Sun-Shin;Wang, Ling;Ryu, Keun-Ho
    • 대한원격탐사학회:학술대회논문집
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    • 대한원격탐사학회 2007년도 Proceedings of ISRS 2007
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    • pp.438-441
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    • 2007
  • Recently, due to molecular biology and engineering technology, DNA microarray makes people watch thousands of genes and the state of variation from the tissue samples of living body. With DNA Microarray, it is possible to construct a genetic group that has similar expression patterns and grasp the progress and variation of gene. This paper practices Cluster Analysis which purposes the discovery of biological subgroup or class by using gene expression information. Hence, the purpose of this paper is to predict a new class which is unknown, open leukaemia data are used for the experiment, and MCL (Markov CLustering) algorithm is applied as an analysis method. The MCL algorithm is based on probability and graph flow theory. MCL simulates random walks on a graph using Markov matrices to determine the transition probabilities among nodes of the graph. If you look at closely to the method, first, MCL algorithm should be applied after getting the distance by using Euclidean distance, then inflation and diagonal factors which are tuning modulus should be tuned, and finally the threshold using the average of each column should be gotten to distinguish one class from another class. Our method has improved the accuracy through using the threshold, namely the average of each column. Our experimental result shows about 70% of accuracy in average compared to the class that is known before. Also, for the comparison evaluation to other algorithm, the proposed method compared to and analyzed SOM (Self-Organizing Map) clustering algorithm which is divided into neural network and hierarchical clustering. The method shows the better result when compared to hierarchical clustering. In further study, it should be studied whether there will be a similar result when the parameter of inflation gotten from our experiment is applied to other gene expression data. We are also trying to make a systematic method to improve the accuracy by regulating the factors mentioned above.

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Transcriptome Analysis of Early Responsive Genes in Rice during Magnaporthe oryzae Infection

  • Wang, Yiming;Kwon, Soon Jae;Wu, Jingni;Choi, Jaeyoung;Lee, Yong-Hwan;Agrawal, Ganesh Kumar;Tamogami, Shigeru;Rakwal, Randeep;Park, Sang-Ryeol;Kim, Beom-Gi;Jung, Ki-Hong;Kang, Kyu Young;Kim, Sang Gon;Kim, Sun Tae
    • The Plant Pathology Journal
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    • 제30권4호
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    • pp.343-354
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    • 2014
  • Rice blast disease caused by Magnaporthe oryzae is one of the most serious diseases of cultivated rice (Oryza sativa L.) in most rice-growing regions of the world. In order to investigate early response genes in rice, we utilized the transcriptome analysis approach using a 300 K tilling microarray to rice leaves infected with compatible and incompatible M. oryzae strains. Prior to the microarray experiment, total RNA was validated by measuring the differential expression of rice defense-related marker genes (chitinase 2, barwin, PBZ1, and PR-10) by RT-PCR, and phytoalexins (sakuranetin and momilactone A) with HPLC. Microarray analysis revealed that 231 genes were up-regulated (>2 fold change, p < 0.05) in the incompatible interaction compared to the compatible one. Highly expressed genes were functionally characterized into metabolic processes and oxidation-reduction categories. The oxidative stress response was induced in both early and later infection stages. Biotic stress overview from MapMan analysis revealed that the phytohormone ethylene as well as signaling molecules jasmonic acid and salicylic acid is important for defense gene regulation. WRKY and Myb transcription factors were also involved in signal transduction processes. Additionally, receptor-like kinases were more likely associated with the defense response, and their expression patterns were validated by RT-PCR. Our results suggest that candidate genes, including receptor-like protein kinases, may play a key role in disease resistance against M. oryzae attack.

혐기성 소화에 미치는 온도와 슬러지의 농도별 고분자 활엽수 리그닌의 영향 (Effects of High Molecular Hardwood Lignin on Anaerobic Digestion at Different Temperatures and Sludge Concentrations)

  • 윤성일;서동일;이성택;김은숙
    • 대한환경공학회지
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    • 제22권12호
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    • pp.2197-2204
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    • 2000
  • 리그닌은 펄프나 제지공장에서 나무의 화학적 처리를 하는 과정에서 생성되는 주요한 부산물이다. 이런 리그닌은 난분해성 물질로서 제지폐수의 생물학적 처리에 어려움을 초래하며, 특히 함량이 높을 경우 혐기성 소화에서 억제(inhibition) 물질로 작용하는 것으로 알려져 있다. 본 연구에서는 회분실험을 통하여 혐기성 소화에서 온도와 혐기성 소화 슬러지의 양에 따른 고분자 활엽수 리그닌(lignosulfonate, MW $\geq$ 20,000)의 영향을 관찰하였다. 고분자 활엽수 리그닌은 혐기성 소화 초기에는 메탄생성에 강한 억제작용을 하였으나 일정한 시간이 지난 후에는 이런 억제작용은 사라지고 메탄생성이 정상적으로 이루어졌다. 즉, 고분자 활엽수 리그닌은 aceticlastic methanogen에 대해 bacteriocidal 작용보다는 bacteriostatic 물질로서 작용하였다. 리그닌이 첨가되지 않은 대조군의 경우에는 메탄생성이 10일간 이루어지는데 반하여 1.3%, 2.6%와 3.9%의 리그닌이 첨가된 경우에는 같은 양의 메탄올 생성하는데 각각 14.5일, 17.8일 과 21.1일이 소요되었다. 2.6%의 리그닌을 첨가한 경우 초기 8일간은 $30^{\circ}C$ 조건에서의 메탄생성속도가 가장 컸으나 12일째부터 $40^{\circ}C$에서의 메탄생성속도가 급속히 증가하여 14일 후에는 총 메탄생성량이 $30^{\circ}C$를 초과하였다. 그러나 $50^{\circ}C$에서는 줄곧 메탄생성이 거의 이루어지지 않았다. 즉, aceticlastic methanogen에 대한 리그닌의 억제작용을 중온 (mesophilic)보다 고온(thermophilic)에서 더 컸다. 리그닌에 의한 이런 억제작용은 또 리그닌의 양(L)과 초기 혐기성 소화슬러지의 농도(AnS)의 비와 중요한 관계가 있었다. L/AnS의 비가 작으면 작을수록 이런 억제작용은 감소되는 것으로 나타났다. 그리고 본 실험에서 고분자 활엽수 리그닌의 분해와 탈색은 이루어지지 않았다.

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Characterization and Partial Nucleotide Sequence Analysis of Alfalfa Mosaic Alfamoviruses Isolated from Potato and Azuki Bean in Korea

  • Jung, Hyo-Won;Jung, Hye-Jin;Yun, Wan-Soo;Kim, Hye-Ja;Hahm, Young-Il;Kim, Kook-Hyung;Choi, Jang-Kyung
    • The Plant Pathology Journal
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    • 제16권5호
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    • pp.269-279
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    • 2000
  • Alfalfa mosaic alfamoviruses(AIMV) were isolated from infected potato (Solanum tuberosum) and azuki bean (Paseolus angularis) in Korea. Two AIMV isolated from potatoes were named as strain KR (AIMV-KR1 and KR2) and AIMV isolated from azuki bean was named as strain Az (AIMV-Az). Each isolated AIMV strain was characterized by using their host ranges, symptom developments, serological relations and nucleotide sequence analysis of coat protein (CP) gene. Strains KR1, KR2, and Az were readily transmitted to 20 of 22 inoculated plant species including bean, cowpea, tomato, tobacco, and potato. AIMV-KR1 and KR2 produced the typical symptoms like chlorotic or necrotic spots in Chenopodium quinoa and Solanum tuberosum cv. Superior. AIMV-Az caused bright yellow mosaic symptom and leaf malformation in Nicotiana glauca, which were different from the common mosaic symptom caused by AIMV-KR1 and KR2. Electron microscope observation of purified virus showed bacilliform virions containing a single-stranded plus-strand RNAs of 3.6, 2.6, 2.0 and 0.9 kbp in length, respectively, similar in size and appearance to those of Alfamovirus. In SDS-PAGE, the coat protein of the two viruses formed a consistent band that estimated to be about 24kDa. The CP genes of the AIMV strains, KR1, KR2, and Az have been amplified by RT-PCR using the specific primers designed to amplify CP gene from viral RNA-3, cloned and sequenced. Computer aided analysis of the amplified cDNA fragment sequence revealed the presence of a single open reading frame capable of encoding 221 amino acids. The nucleotide and peptide sequence of viral CP gene showed that strain KR1, KR2, and Az shared highest nucleotide sequence identities with AIMV strain 425-M at 97.7%, 98.2%, and 97.2%, respectively. CP gene sequences of two strains were almost identical compared with each other. Altogether, physical, serological, biological and molecular properties of the purified virus.

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