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한국해안으로부터 Purple, Non-Sulfur Photosynthetic Bacterium, Rhodobacter sp. EGH-24의 분리 및 특성
차미선,김기한,조순자,이나은,이정은,이재동,이상준,박재림 한국환경과학회 2003 한국환경과학회지 Vol.12 No.12
A species of facultative photo-organotrophic, purple, non-sulfur bacterium was isolated from the 47 point at west and south coast of Korea in September 2001. Separated 13 samples of changes with red color under 28~32 ℃, 3000 lux, anaerobe conditions for 7 days cultivated in basal medium. For pure isolation from 13 samples, we used agar-shake tube method (0.4 % agar) and separated 5 strains through 13-repetition test. EGH-24 and EGH-30 was identified as the same strain through the RAPD(Random Amplified Polymorphic DNA)-PCR of strain EGH-9, EGH-13, EGH-23, EGH-24, EGH-30. Four isolates cultivated in synthesis wastewater for wastewater biodegradation test. EGH-24 was selected with efficient wastwater treating strain. Based on the results obtained from morphology, nutrient requirements, major bacteriochlorophyll content, 16S-rDNA phylogenetic analysis, EGH-24 strain may be identified as a new strain of the genus Rhodobacter and named Rhodobacter sp. EGH-24.
Jung, Hee-Young,Kang, Seung-Mi,Kang, Young-Min,Kang, Min-Jung,Yun, Dea-Jin,Bahk, Jung-Dong,Yang, Jae-Kyung,Choi, Myung-Suk Plant molecular biology and biotechnology research 2003 Plant molecular biology and biotechnology research Vol.2003 No.-
In an attempt to increase productivity, the effect of elicitation on tropane alkaloids (TA) biosynthesis was studied in adventitious hairy root cultures of Schopolia parviflora. Two Gram-positive strains and one Gram-negative strain of bacteria were used as biotic elicitors. The raw bacterial elicitors affected the tropane alkaloid profile by increasing the scopolamine concentration, while the autoclaved bacterial elicitors produced similar effects on the control. The conversion ratio of hyoscyamine to scopolamine was increased following elicitation using raw bacteral elicitors. The bacteral elicitor inhibited the expression of H6H (hyoscyamine 6β-hydoxylase) whereas the expression of PMT (putrescine N-methyltransferase) was rainsed by elicitation. These results have important implications for the large-scale production of tropane alkaloids.
Proteomic analysis of apoptosis related proteins regulated by proto-oncogene protein DEK
Kim, Dong-Wook,Chae, Jung-IL,Kim, Ji-Young,Pak, Jhang Ho,Koo, Deog-Bon,Bahk, Young Yil,Seo, Sang-Beom Wiley Subscription Services, Inc., A Wiley Company 2009 Journal of cellular biochemistry Vol.106 No.6
<P>A nuclear phosphoprotein, DEK, is implicated in certain human diseases, such as leukemia and antoimmune disorders, and a major component of metazoan chromatin. Basically as a modulator of chromatin structure, it can involve in various DNA and RNA-dependent processes and function as either an activator or repressor. Despite of numerous efforts to suggest the biological role of DEK, direct target proteins of DEK in different physiological status remains elusive. To investigate if DEK protein triggers the changes in certain protein networks, DEK was knocked down at both types of cell clones using siRNA expression. Here we provide a catalogue of proteome profiles in total cell lysates derived from normal HeLa and DEK knock-down HeLa cells and a good in vitro model system for dissecting the protein networks due to this proto-oncogenic DEK protein. In this biological context, we compared total proteome changes by the combined methods of two-dimensional gel electrophoresis, quantitative image analysis and MALDI-TOF MS analysis. There were a large number of targets for DEK, which were differentially expressed in DEK knock-down cells and consisted of 58 proteins (41 up-regulated and 17 down-regulated) differentially regulated expression was further confirmed for some subsets of candidates by Western blot analysis using specific antibodies. In the identified 58 spots, 16% of proteins are known to be associated with apoptosis. Among others, we identified apoptosis related proteins such as Annexins, Enolase1, Lamin A, and Glutathione-S-transferase omega 1. These results are consistent with recent studies indicating the crucial role of DEK in apoptosis pathway. We further demonstrated by ChIP analysis that knock-down of DEK caused hyperacetylation of histones around Prx VI promoter which is upregulated in our profile. Using immunoblotting analysis, we have demonstrated the modulation of other caspase-dependent apoptosis related proteins by DEK knock-down and further implicate its role in apoptosis pathway. J. Cell. Biochem. 106: 1048–1059, 2009. © 2009 Wiley-Liss, Inc.</P>
Molecular and functional characterization of a PEX14 cDNA from rice
Lee, Jung-Ro,Lee, Kyun-Oh,Park, Jin-Ho,Yoo, Ji-Young,Kang, Jae-Sook,Jeon, Hye-Sook,Kim, Sun-Young,Lee, Young-Mi,Kim, Sun-Tae,Lim, Chae-Oh,Bahk, Jeong-Dong,Cho, Moo-Je,Lee, Sang-Yeol Plant molecular biology and biotechnology research 2003 Plant molecular biology and biotechnology research Vol.2003 No.-
In contrast to the translocation mechanisms determined in yeasts and mammalian cells, there is little information on the functions of plant peroxisomal proteins or their genomic structures. To understand the role that PEX14 plays in diverse plant peroxisomal functions and how peroxisomal translocation is mediated in plant cells, we cloned a 1827 bp cDNA encoding the peroxisomal membrane protein OsPex14p from a rice leaf cDNA library. The 54kDa OsPex14p, which has a theoretical pI value of 6.06, contains a highly conserved N-terminal domain and a short putative transmembrane domain. The OsPEX14 gene in the rice genome exists as a single-copy gene, consists of eleven exons interrupted by ten introns, and spans about 5kb of chromosome 5. The 5′-flanking region contains putative cis-acting light-responsive elements, and the transcription initiation site maps 114bp upstream of the translation start codon. OsPEX14 mRNA is highly expresssed in leaf tissues and is induced by exposure to several stresses. Heterologous expression of OsPex14p suppresses the defect in targeting of peroxisomal matrix proteins in a pex14 null mutant of Saccharomyces cerevisiae.
A proteomic approach in analyzing heat-responsive proteins in rice leaves
Lee, Dong-Gi,Ahsan, Nagib,Lee, Sang-Hoon,Kang, Kyu Young,Bahk, Jeong Dong,Lee, In-Jung,Lee, Byung-Hyun WILEY-VCH 2007 Proteomics Vol. No.
<P>The present study investigated rice leaf proteome in response to heat stress. Rice seedlings were subjected to a temperature of 42°C and samples were collected 12 and 24 h after treatment. Increased relative ion leakage and lipid peroxidation suggested that oxidative stress frequently was generated in rice leaves exposed to high temperature. 2-DE, coupled with MS, was used to investigate and identify heat-responsive proteins in rice leaves. In order to identify the low-abundant proteins in leaves, samples were prefractionated by 15% PEG. The PEG supernatant and the pellet fraction samples were separated by 2-DE, and visualized by silver or CBB staining. Approximately 1000 protein spots were reproducibly detected on each gel, wherein 73 protein spots were differentially expressed at least at one time point. Of these differentially expressed proteins, a total of 34 and 39 protein spots were found in the PEG supernatant and pellet fractions, respectively. Using MALDI-TOF MS, a total of 48 proteins were identified. These proteins were categorized into classes related to heat shock proteins, energy and metabolism, redox homeostasis, and regulatory proteins. The results of the present study show that a group of low molecular small heat shock proteins (sHSPs) were newly induced by heat stress. Among these sHSPs, a low molecular weight mitochondrial (Mt) sHSP was validated further by Western blot analysis. Furthermore, four differentially accumulated proteins that correspond to antioxidant enzymes were analyzed at the mRNA level, which confirmed the differential gene expression levels, and revealed that transcription levels were not completely concomitant with translation. The identification of some novel proteins in the heat stress response provides new insights that can lead to a better understanding of the molecular basis of heat-sensitivity in plants.</P>
Park, Soo-Kwon,Lee, Jung-Ro,Lee, Seung-Sik,Son, Hyo-Jin,Yoo, Ji-Young,Moon, Jeong-Chan,Kwon, Heun-Young,Lim, Chae-Oh,Bahk, Jeong-Dong,Cho, Moo-Je,Lee, Sang-Yeol Plant molecular biology and biotechnology research 2002 Plant molecular biology and biotechnology research Vol.2002 No.-
Three soluble enzyme fractions (F-Ⅰ, F-Ⅱand F-Ⅲ)that hydrolyze phophoinositides were separated from soybean sprouts by using Matrex green gel column chromatography. Among the three phosphatidylinositol (PI)-specific phopholipsase C (PLC) enzymes, only the third fraction (F-Ⅲ) was able to hydrolyze phosphatidylinositol 4,5-bisphosphate (PIP_(2)) as well as phosphatidylinositol (PI) and phosphatidylinositol phosphate(PIP) as substrates. The F-Ⅰand F-Ⅱ fraction only showed enzymatic activities for PI and PIP. The PIP_(2)-hydrolyzing PLC protein, F-Ⅲ, was partially purified using the chromatographic steps of the Matrex green gel, phenyl Toyopearl, Matrex orange gel, Mono S cation exchange, and superose 6 gel filtration columns. The molecular weight of the F-Ⅲ protein was estimated to be about 64 kDa on SDS-PAGE. The protein showed immunocross-reactivity with a polyclonal antibody that was prepared against the X and Y motifs of animal PLC enzymes, the conserved catalytic domains. Ca^(2+) ion critically affected the PIP_(2)-hydrolyzing PLC activity of F-Ⅲ protein, representing maximal activity at 10㎛ Ca^(2+) concentration. The PIP_(2)-hydrolyzing PLC activity of the protein was also significantly increased by sodium deoxycholate (SDC) from 0.05 to 0.08%. However, the activity was greatly reduced above the concentration, and no activity was detected at 0.3% SDC. In addition, the protein exhibited maximal PIP_(2)-hydrolyzing PLC activity at pH, in the range of 6.5-7.5.
Kang, Seung-Mi,Jung, Hee-Young,Kang, Young-Min,Yun, Dae-Jin,Bahk, Jung-Dong,Yang, Jae-kyung,Choi, Myung-Suk Plant molecular biology and biotechnology research 2004 Plant molecular biology and biotechnology research Vol.2004 No.-
The effects of methyl jasmonate (MJ) and salicylic acid (SA) on the production of tropane alkaloids (TA) and the expression of putrescine N-methy1transferase (PMT) and hyoscyamine 6β-hydroxylase (H6H) were studied in adventitious root cultures of Scopolia parviflora. MJ treatments increased the amounts of both scopolamine and hyoscyamine, with growth inhibition of the roots, while SA increased the amount of scopolamine without negative effects on growth. Regarding both MJ and SA, their concentrations and exposure times were factors that strongly affected the production of TA. Western blot analysis showed that treatments with MJ or SA regulated the expression of the key enzymes PMT and H6H gene and, finally, increased the production of TA and stimulated the excretion of TA into the culture medium.