• Title/Summary/Keyword: 2-D PAGE

Search Result 323, Processing Time 0.035 seconds

Induction of Drought Stress Resistance by Multi-Functional PGPR Bacillus licheniformis K11 in Pepper

  • Lim, Jong-Hui;Kim, Sang-Dal
    • The Plant Pathology Journal
    • /
    • v.29 no.2
    • /
    • pp.201-208
    • /
    • 2013
  • Drought stress is one of the major yield affecting factor for pepper plant. The effects of PGPRs were analyzed in relation with drought resistance. The PGPRs inoculated pepper plants tolerate the drought stress and survived as compared to non-inoculated pepper plants that died after 15 days of drought stress. Variations in protein and RNA accumulation patterns of inoculated and non-inoculated pepper plants subjected to drought conditions for 10 days were confirmed by two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and differential display PCR (DD-PCR), respectively. A total of six differentially expressed stress proteins were identified in the treated pepper plants by 2D-PAGE. Among the stress proteins, specific genes of Cadhn, VA, sHSP and CaPR-10 showed more than a 1.5-fold expressed in amount in B. licheniformis K11-treated drought pepper compared to untreated drought pepper. The changes in proteins and gene expression patterns were attributed to the B. licheniformis K11. Accordingly, auxin and ACC deaminase producing PGPR B. licheniformis K11 could reduce drought stress in drought affected regions without the need for overusing agrochemicals and chemical fertilizer. These results will contribute to the development of a microbial agent for organic farming by PGPR.

Purification and Properties of D-Xylose Isomerase from Lactococcus sp. JK-8 (Lactococcus sp. JK-8에서 생산된 D-Xylose isomerase의 정제와 특성에 관한 연구)

  • Jun, Hong-Ki;Kim, Suk-Young;Baik, Hyung-Suk
    • Journal of Life Science
    • /
    • v.14 no.4
    • /
    • pp.636-643
    • /
    • 2004
  • D-Xylose isomerase produced by Lactococcus sp. JK-8, isolated from kimchi, was purified 17-fold of homogeneity, and its physicochemical properties were determined. Although the N-terminal amino acid sequence of D-xylose isomerase was analysed to Ala-Tyr-Phe-Asn-Asp-Ile-Ala-Pro-Ile-Lys, it was not similar to that of Lactobacillus enzyme. The molecular weight of the purified enzyme was estimated to be 180 kDa by gel filtration, 45 kDa by SDS-PAGE and the enzyme was homotetramer. The optimum pH of the enzyme was around 7 and stable between pH 6 and 8. The optimum reaction temperature was 7$0^{\circ}C$ and stable up to 7$0^{\circ}C$ in the presence of 1 mM $Mn^{2+}$. Like other D-xylose isomerases, this enzyme required divalent cation, such as $Mg^{2+}$, $Co^{2+}$, or $Mn^{2+}$ for the activity and thermostability. $Mn^{2+}$was the best activator. Substrate specificity studies showed that this enzyme was highly active on D-xylose. The enzyme had an isoelectric point of 4.8, and fm values for D-xylose was 5.9 mM.

Difference in Electrophoretic Phenotypes of rice Cultivars Selected to Bensulfuron (Bensulfuron에 대(對)한 내성(耐性) 및 감수성(感受性) 수도품종(水稻品種)의 전기영동(電氣泳動) 표현형(表現型) 차이(差異))

  • Kuk, Y.I.;Guh, J.O.;Kim, Y.J.;Lee, D.J.
    • Korean Journal of Weed Science
    • /
    • v.8 no.3
    • /
    • pp.250-257
    • /
    • 1988
  • The study was intended to know any relations between the rice tolerance to bensulfuron and varietal speciation in seed protein composition or any enzymatical allelies with or without chemical treatment. Rice varieties used were UCP-28, Chinsurah Boro II, Fukunohama, Fadehpur-2, IR 14252-13-2-2-5 as the tolerant group, and HP 93(3) FA, HP94(9) FA, Padilabou Alumbis, KH-17854, and IR 1846-2841-1 as the susceptible, respectively. Electrophoretic methods used were SDS-PAGE for seed protein, 7% PAGE for isozymes (acid phosphatase, peroxidase, malate dehydrogenase, and esterase from rice seedling) and variation in isoenzyme profiles (malate dehydrogenase, peroxidase, and esterase) as affected by different concentrations of bensulfuron(0, $10^{-6}$, $10^{-5}$ and $3{\times}10^{-5}M$) was also studied. The results are summarized as follows. -Among 16 bands separated in seed proteins, two different rice groups selected in terms of tolerance to bensulfuron were clustered in dissimilarity, which was based on relatively larger area in whole peaks and higher activities in N, O, P bands for the tolerant group. -Among isozymes obtained from rice seedlings without chemical treatments, the following specificities were obtained. The tolerant varieties had the relatively higher activity in D band out of 4 peroxidase bands. Malate dehydrogenase was separated into 3 bands and only tolerant varieties had A band and higher activities in Band C bands. Esterase was separated into 3-4 bands with higher activities in A and B bands for tolerant varieties. There were one major band accompanied by 2-3 minor bands for acid phosphatase in which only tolerant varieties had the B band. -The effect of Bensulfuron concentration on the isozyme activities showed that the activity of C band in peroxidase was not present in tolerant varieties which was contrary to the increased activities in susceptible varieties. However, D band was gradually disappeared only in susceptible varieties as the concentration of bensulfuron was increased. For malate dehydrogenase in the susceptible varieties, major bands D, E and F kept consistantly higher activities while minor bands A, B and C disappeared sensitively. Among 5 bands of esterase separated, D band was present only in the tolerant varieties while E band only in the susceptible. The activities in A, C, E bands were sharply decreased in the susceptible varieties as the concentration of bensulfuron was increased.

  • PDF

Disassembly of Chlorophyll-Protein Complexes in Arabidopsis thaliana during Dark-Induced Foliar Senescence

  • Lee, Choon-Hwan
    • Journal of Plant Biology
    • /
    • v.39 no.4
    • /
    • pp.301-307
    • /
    • 1996
  • The disassembly of Chl-protein complexes during dark-induced senescence (DIS) was investigated using detached third and fourthleaves of 21$\pm$1 day-old Arabidopsis thaliana. Although Chl content decreased linearly after 1 d, a significant decrease of photochemical effeciency (Fv/Fm) was observed after 2 d. In experiments using native green gel electrophoresis of Chl-protein complexes combined with additional two-dimensional SDS-PAGE analysis, we could observe the degradation of both photosystems after 2 d. Although light-harvesting complex(LHC) for PSI (LHCI) was degraded first in PSI complex, small PSII apoproteins including CP47/CP43 and D1/D2 apoproteins were degraded first in PSII complexes. LHC for PSII (LHCII) trimers were stable until 4 d. The level of LHCII monomers was increased until 3 and decreased thereafter, resulting in the increase of free pigments. These results suggest that the disassembly process of PSI is different from that of PSII.

  • PDF

Effects of Several Medicinal Plants Extract on Survival Rate, Chlorophyll Contents and Photosynthetic Electron Transport Activity of Liverwort Photoautotrophic Cultured Cell (약용식물 추출액이 우산이끼 자가관양배양세포의 생존율, 엽록소함량 및 광합성전자전달 활성에 미치는 영향)

  • 정형진;권순태;김시무
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.40 no.2
    • /
    • pp.133-141
    • /
    • 1995
  • The effects of allelochemicals from medicinal plants have been studied as photo-synthetic inhibitor for photoautotrophic(PA) cultured cells. The extracts from 9 plant species were used for measuring the physiological effects on the liverwort cultured cell in following areas; germination inhibition, chlorophyll contents, hill activity, cell viability, photosynthetic oxygen evolution,and protein pattern changes on SDS PAGE. Germination inhibitions were detected in all plant after treating with 10% extract. Especially, treatment with 10% extract from Pulsatilla koreana and Aconitum carmichael inhibited germinations completely. Chlorophyll fornation was inhibited completely by treating PA cells with extract of Pulsatilla koreana, whose effect was similar to that of DCMU 10-3M, inhibitor for photosynthetic electron trans-fer. The treatment with extract from Pulsatilla koreana on PA cell showed the highest hill activity and the lowest cell viability among extracts studied. Oxygen releasing has been decreased down to 14-77% after treating with extracts from Pinellia ternata, Araliacont inentaila, Pulsatilla koreana and Vitex rotundifolia. Especially, 60$\mu$l of Pulsatilla koreana extract into 2ml mixture of PA cell inhibit-ed oxygen release up to 50%. Protein bands on SDS-PAGE, 14kD, 31kD, 41kD, 53kD, and 73kD, were not detected after treating Pulsatilla koreana extract on PA cells.

  • PDF

Comparative Proteomic Analysis of Virulent Korean Mycobacterium tuberculosis K-strain with Other Mycobacteria Strain Following Infection of U-937 Macrophage

  • Ryoo, Sung-Weon;Park, Young-Kil;Park, Sue-Nie;Shim, Young-Soo;Liew, Hyun-Jeong;Kang, Seong-Man;Bai, Gill-Han
    • Journal of Microbiology
    • /
    • v.45 no.3
    • /
    • pp.268-271
    • /
    • 2007
  • In Korea, the Mycobacterium tuberculosis K-strain is the most prevalent clinical isolates and belongs to the Beijing family. In this study, we conducted comparative porteomics of expressed proteins of clinical isolates of the K-strain with H37Rv, H37Ra as well as the vaccine strain of Mycobacterium bovis BCG following phagocytosis by the human monocytic cell line U-937. Proteins were analyzed by 2-D PAGE and MALDI-TOF-MS. Two proteins, Mb1363 (probable glycogen phosphorylase GlgP) and MT2656 (Haloalkane dehalogenase LinB) were most abundant after phagocytosis of M. tuberculosis K-strain. This approach provides a method to determine specific proteins that may have critical roles in tuberculosis pathogenesis.

The Impacts of Photoperiods on Hypothalamic Proteins in the Reproductive Activities of Golden Hamsters

  • Choi, Don-Chan;Han, Eun-Hee
    • Development and Reproduction
    • /
    • v.14 no.3
    • /
    • pp.185-197
    • /
    • 2010
  • The reproduction of animals is a means to maintain their species. The golden hamsters are seasonal breeders, and their reproduction is regulated by the photoperiod. The shifts of the sexual functions imply the changes of the protein expression, and those are reflected by the proteome. Thus the present study was to examine via two dimensional polyacrylamide gel electrophoresis (2d PAGE), the physiological changes and the alterations of protein expressions in hypothalamus upon the reproductive situation related to the pineal gland. Among the pineal intact animals, the reproductive functions were sustained in animals housed in long photoperiod (LP) but regressed in animals housed in short photoperiod. And those pinealectomized animals showed high sexual activities regardless of photoperiod. Ultimately they were branched into dichotomy, sexually active and inactive animals. Apart from the changes of physiological parameters upon the reproductive conditions, there were obvious differences in proteins extracted from the hypothalamus. The hypothalamus of LP animals presented high levels of enzymes which are involved in the production of energy, glycolysis and Krebs cycle. The increased energy might be related to the GnRH synthesis in hypothalamus and indirectly to the constant cell divisions in spermatogenesis. Taken together, the impacts of the photoperiodic information controlling reproduction could be observed through 2d PAGE. Therefore, the present results suggest the potential of biomarkers collectively to diagnose the fertility and the infertility by way of proteomics in organs with regard to the reproductive system, further could be applied to diagnose various diseases.

An Evaluation of Changes in the Allergenicity of Kochujang upon Preparation Using Aloe Extract

  • Son, Bo-Kyung;Huh, Yoon-Ee;Kim, Jung-Yun;Noh, Geon-Woong;Lee, Sang-Sun
    • Nutritional Sciences
    • /
    • v.9 no.4
    • /
    • pp.317-322
    • /
    • 2006
  • Soybeans are well-known as allergenic foods. Koreans consume large amounts of soybean foods, such as kochujang, which have gone through the fermentation process. To lower the allergenicity of these foods, we prepared hypo allergenic kochujang with aloe extract (AK). A sensory evaluation was conducted along with a clinical evaluation that used a double-blind, placebo-controlled food challenge (DBPCFC) test These tests were designed to evaluate the acceptability of the fermented foods. In comparison to normal kochujang (NK), AK elicited a higher sensory test score, and the rate of positive reactions in atopic dermatitis patients during the DBPCFC test was reduced. Methods of protein extraction, protein quantitation with sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), and protein identification using two-dimensional (2D) gel electrophoresis were performed for both NK and AK to compare the functional factors. We found a reduction in the levels of high molecular proteins even though the bands of the proteins had not entirely disappeared, indicating that the boiling and fermentation process changed the soybean protein patterns. The rate of the reduction of high molecular proteins was more effective in the AK. In conclusion, AK can be recognized as a food with hypoallergenic effect.

Cellular Responses of the TNT-degrading Bacterium, Stenotrophomonas sp. OK-5 to Explosive 2,4,6-Trinitrotoluene (TNT) (폭약 2,4,6-Trinitrotoluene에 노출된 분해세균 Stenotrophomonas sp. OK-5의 세포반응)

  • 장효원;송승열;김승일;강형일;오계헌*
    • Korean Journal of Microbiology
    • /
    • v.38 no.4
    • /
    • pp.247-253
    • /
    • 2002
  • The cellular responses of TNT-degrading bacterium, Stenotrophomonas sp. OK-5 to explosive 2,4,6-trini-trotoluene (TNT) as an environmental contaminant were examined. Survival of the strain OK-5 with time in the presence of different concentrations of TNT under sublethal conditions was monitored, and viable counts paralleled the production of the stress shock proteins in this bacterium. Total cellular fatty acids analysis showed that strain OK-5 produced or disappeared several different kinds of lipids when grown on TNT media than when grown on TSA. Under scanning electron microscope, the cells treated with 0.5 mM TNT for 12 hrs showed irregular rod shapes with wrinkled surfaces. Analyses of SDS-PAGE and Western blot using anti-DnaK and anti-GroEL revealed that several stress shock proteins including 70 kDa DnaK and 60 kDa GroEL in strain OK-5 were newly synthesized at different TNT concentrations in exponentially growing cultures. 2-D PAGE of soluble protein fractions from the culture of OK-5 exposed to TNT demonstrated that approximately 300 spots were observed on the silver stained gel ranging from pH 3 to pH 10. Among them, 10 spots significantly induced and expressed in response to TNT were selected and analyzed. As the result of internal amino acid sequencing with ESI-Q TOF, two proteins, spot #1 and spot #10 were assigned the DnaK protein XF2340 of Xylella fastidiosa and stress-induced protein of Mesorhizobium loti, respectively.

Prefetch R-tree: A Disk and Cache Optimized Multidimensional Index Structure (Prefetch R-tree: 디스크와 CPU 캐시에 최적화된 다차원 색인 구조)

  • Park Myung-Sun
    • The KIPS Transactions:PartD
    • /
    • v.13D no.4 s.107
    • /
    • pp.463-476
    • /
    • 2006
  • R-trees have been traditionally optimized for the I/O performance with the disk page as the tree node. Recently, researchers have proposed cache-conscious variations of R-trees optimized for the CPU cache performance in main memory environments, where the node size is several cache lines wide and more entries are packed in a node by compressing MBR keys. However, because there is a big difference between the node sizes of two types of R-trees, disk-optimized R-trees show poor cache performance while cache-optimized R-trees exhibit poor disk performance. In this paper, we propose a cache and disk optimized R-tree, called the PR-tree (Prefetching R-tree). For the cache performance, the node size of the PR-tree is wider than a cache line, and the prefetch instruction is used to reduce the number of cache misses. For the I/O performance, the nodes of the PR-tree are fitted into one disk page. We represent the detailed analysis of cache misses for range queries, and enumerate all the reasonable in-page leaf and nonleaf node sizes, and heights of in-page trees to figure out tree parameters for best cache and I/O performance. The PR-tree that we propose achieves better cache performance than the disk-optimized R-tree: a factor of 3.5-15.1 improvement for one-by-one insertions, 6.5-15.1 improvement for deletions, 1.3-1.9 improvement for range queries, and 2.7-9.7 improvement for k-nearest neighbor queries. All experimental results do not show notable declines of the I/O performance.