• Title/Summary/Keyword: Heat-shock protein

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Analysis of Expressed Sequence Tags from the Embryogenic Callus of Korean Ginseng (Panax ginseng C.A. Meyer)

  • In, Jun-Gyo;Lee, Bum-Soo;Park, Yong-Eui;Yang, Deok-Chun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.123-123
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    • 2003
  • In order to study gene expression transcribted during the embryo development, we constructed a cDNA library of embryogenic callus induced from cotylendon of Korean ginseng and generated expressed sequence tags (ESTs) of 3,359 clones randomly selected. The ESTs could be clustered into 1,910 (59.1%) non-redundant groups. Similarity search of the non-redundant ESTs against public non-redundant databases of both protein and DNA indicated that 2,217 groups show similarity to genes of known function. These ESTs clones were divided into eighteen categories depending upon gene function. Most abundant transcripts were ribosomal protein small subunit 28kDa(40), tumor-related protein(35), metallothionein (31), small heat-shock protein class 18.6K(24), and cyclophilin(20). There are no useful informations of gene expression during the embryo development in Korean ginseng. These results could help to understand the embryo development in Korean ginseng.

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Control Effect of a Stored Grain Insect Pest, Tribolium castaneum, by 'CATTS' Postharvest Treatment (CATTS를 이용한 저곡해충 거짓쌀도둑거저리(Tribolium castaneum)의 소독 효과)

  • Son, Ye-Rim;Kim, Yong;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.49 no.4
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    • pp.363-369
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    • 2010
  • A postharvest treatment called CATTS (controlled atmosphere and temperature treatment system) has been used as an alternative nonchemical measure for methyl bromide fumigant treatment. This study applied CATTS to control the red flour beetle, Tribolium castaneum, infesting stored grains. Adults of T. castaneum were susceptible to $46^{\circ}C$ heat treatment. The susceptibility was further enhanced by addition of CA conditions (15% $CO_2$ and 1% $O_2$). When CATTS ($46^{\circ}C$, 15% $CO_2$, $16^{\circ}C/h$ treating rate) was applied to different developmental stages of T. castaneum, it showed 100% control efficacy by 120 min exposure. There was a variation in CATTS susceptibility among developmental stages, in which late instar larvae were most tolerant. Heat shock proteins of T. castaneum appeared to be implicated in the tolerance of CATTS.

Overexpression of NtHSP70-1 Protects Chlorophyll from High Temperature in Plants (NtHSP70-1에 의한 클로로필의 고온 내성 효과)

  • Cho, Eun-Kyung;Hong, Choo-Bong
    • Journal of Life Science
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    • v.18 no.3
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    • pp.304-310
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    • 2008
  • Heat shock protein 70 (HSP70) is known as molecular chaperone, the fundamental protein participating in various processes, from nascent protein synthesis to protection of proteins during abiotic stresses and developmental programs. However, their biological functions in plants are not yet well known. Here, NtHSP70-1 (AY372069), HSP70 of Nicotiana tabacum induced by heat stress was investigated. To analyze the protective role of NtHSP70-1, transgenic tobacco plants, which constitutively overexpressed NtHSP70-1 as well as contained either the vector alone or having NtHSP70-1 in the antisense orientation, were constructed. The altered NtHSP70-1 levels in plants were confirmed by western blotting and transgenic sense lines exhibited tolerance to heat stress. Seedlings with the constitutively expressed NtHSP70-1 grew as green or healthy plants after heat stress. In contrast, transgenic vector or antisense lines exhibited yellowing of leaves or some delay in growth, which finally led to death. Evaluation of chlorophyll contents of heat-shocked transgenic tobacco seedlings indicated that NtHSP70-1 contributes to thermotolerance by preventing chlorophyll synthesis in plants.

Intratumoral Administration of Dendritic Cells Combined with Hyperthermia Induces Both Local and Systemic Antitumor Effect in Murine Tumor Models (온열 요법 후 종양 내 주입한 수지상 세포의 국소 및 원격 항종양 효과)

  • Kwon Byung-Hyun;Kim Won-Taek;Kim Young-Kan;Kim Dong-Won
    • Radiation Oncology Journal
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    • v.24 no.1
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    • pp.51-57
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    • 2006
  • Puroose: We examined whether intratumoral (i.t.) administration of dendritic cells (DCs) into a treated tumor could induce local and systemic antitumor effects in a mouse tumor model. Methods and Materials: C57BL/6 mice were inoculated s.c. in the right and left thighs with MCA-102 fibrosarcoma cells on day 0 and on day 7, respectively. On day 7, the tumors (usually 6 mm in diameter) on the right thigh were heated by immersing the tumor-bearing leg in a circulating water bath at $43^{\circ}C$ for 30 min; thereafter, the immature DCs were i.t administered to the right thigh tumors. This immunization procedure was repeated on days 7, 14 and 21. The tumors in both the right and left thighs were measured every 7 days and the average sizes were determined by applying the following formula, tumor $size=0.5{\times}(length+width)$. Cytotoxicity assay was done to determine tumor-specific cytotoxic T-lymphocyte activity. Results: Hyperthermia induced apoptosis and heat shock proteins (HSPs) in tumor occurred maximally after 6 hr. For the local treated tumor, hyperthermia (HT) alone inhibited tumor growth compared with the untreated tumors (p<0.05), and furthermore, the i.t. administered DCs combined with hyperthermia (HT + DCs) additively inhibited tumor growth compared with HT alone (p<0.05). On the distant untreated tumor, HT alone significantly inhibited tumor growth (p<0.05), and also HT + DCs potently inhibited tumor growth (p<0.001); however, compared with HT alone, the difference was not statistically significant. In addition, HT + DCs induced strong cytotoxicity of the splenocytes against tumor cells compared to DCs or HT alone. Conclusion: HT + DCs induced apoptosis and increased the expression of HSPs, and so this induced a potent local and systemic antitumor response in tumor-bearing mice. This regimen may be beneficial for the treatment of human cancers.

Identification of Highly Transcribed Genes in Japanese Oak Silkworm, Antheraea yamamai, Using PCR-Based cDNA Library

  • Lee, Jin-Sung;Kim, Ki-Hwan;Goo, Tae-Won;Yun, Eun-Young;Kang, Seok-Woo;Suh, Dongs-Sang;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.1 no.2
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    • pp.171-175
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    • 2000
  • Determined sequences of 384 randomly selected clones in a PCR-based cDNA library of Antheraea yamamai could identify expressed sequence tags (ESTs) of highly expressed gene. One EST (fibroin) appeared 15 times, one EST (40S ribosomal protein S18) twelve times, one EST (ribosomal protein S24a) eleven times, ten times (ribosomal protein S8), nine times (60S ribosomal protein L10A), seven times (60S ribosomal protein S15A, S17, S17 and seroin), six times (ribosomal protein S8), five times (ribosomal protein S24, mariner transposase and P8 protein), four times (serpin 2), three times (heat shock protein 70 and poly A binding protein), and the remaining 6 ESTs twice (amylase, KIAA1006, elongation factor-1, transposon mag, translation initiation factor 4C, QM protein, transposase). Therefore, the 94 EST make it possible to identify 24 redundant clones that are candidates for highly expressed genes in posterior silk gland of this insect. The 24 redundant EST clones were identified in GenBank, but none of them was related to A. yamamai, suggesting that there are many unidentified genes which are highly expressed in the A. yamamai genome.

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Molecular Characterization of Ischemia-Responsive Protein 94 (irp94) Response to Unfolded Protein Responses in the Neuron

  • Kim Seung-Whan;Kwon Ki-Sang;Shin Kee-Sun;Kim Seung-Ho;Kwon O-Yu
    • Biomedical Science Letters
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    • v.12 no.2
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    • pp.81-89
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    • 2006
  • The ischemia-responsive 94 gene (irp94) encoding a 94 kDa endoplasmic reticulum resident protein was investigated its molecular properties associated with unfoled protein responses. First, the expression of irp94 mRNA was tested after the reperfusion of the transient forebrain ischemia induction at the central nervous system in three Mongolian gerbils. Second, irp94 expression in PC12 cells, which are derived from transplantable rat pheochromocytoma cultured in the DMEM media, was tested at transcriptional and translational levels. The half life of irp94 mRNA was also determined In PC12 cells. Last, the changes of irp94 mRNA expression were investigated by the addition of various ER stress inducible chemicals (A23187, BFA, tunicamycin, DTT and $H_2O_2$) and proteasome inhibitors, and heat shock. High level expression of irp94 mRNA was detected after 3 hours reperfusion in the both sites of the cerebral cortex and hippocampus of the gerbil brain. The main regulation of irp94 mRNA expression in PC 12 cells was determined at the transcriptional level. The half life of irp94 mRNA in PC12 cells was approximately 5 hours after the initial translation. The remarkable expression of irp94 mRNA was detected by the treatment of tunicamycin, which blocks glycosylation of newly synthesized polypeptides, and $H_2O_2$, which induces apoptosis. When PC12 cells were treated with the cytosol proteasome inhibitors such as ALLN (N-acetyl-leucyl-norleucinal) and MG 132 (methylguanidine), irp94 mRNA expression was increased. These results indicate that expression of irp94 was induced by ER stress including oxidation condition and glycosylation blocking in proteins. Expression of irp94 was increased when the cells were chased after heat shock, suggesting that irp94 may be involved in recovery rather than protection against ER stresses. In addition, irp94 expression was remarkably increased when cytosol proteasomes were inhibited by ALLN and MG 132, suggesting that irp94 plays an important role for maintaining the ERAD (endoplasmic reticulum associated degradation) function.

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Relationship between Plasminogen Activator (PA) and HSP-90 in Uterus Tissue during the Porcine Estrous Cycle (돼지의 발정 주기 동안 자궁조직에서 Plasminogen Activator(PA)와 HSP-90과의 관계)

  • Gu, Ha-Na;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.219-223
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    • 2012
  • The present study was performed to identify the relationship between plasminogen activator (PA) and Heat Shock Protein-90 (HSP-90) in porcine uterus tissues during the estrous cycle. Porcine uterus tissues were obtained from preovulatory (Pre-Ov), post-ovulatory (Post-Ov) and early to mid-luteal (Early-mid L) stages. The protein was extracted from uterus tissue by using M-PER Mammalian Protein Extraction Reagent. Proteins were refined by RIPA Buffer and quantified by BCA methods. As results, t-PA expression was significantly (p<0.05) higher from pre-ovulatory(Epithelium tissue: $29,067{\mu}g/{\mu}l$, Myometrium tissue: $30,797{\mu}g/{\mu}l$) compared to the post-ovulatory stage(Epithelium tissue: $54,357{\mu}g/{\mu}l$, Myometrium tissue: $53,270{\mu}g/{\mu}l$) and early to mid-luteal stage(Epithelium tissue: $42,380{\mu}g/{\mu}l$, Myometrium tissue: $43,139{\mu}g/{\mu}l$). On the other hand, the uPA expression indicated higher from early to mid-luteal stage (Epithelium tissue: $0.02198{\mu}g/{\mu}l$, Myometrium tissue: $0.02412{\mu}g/{\mu}l$) than pre-ovulatory stage (Epithelium tissue: $0.01577{\mu}g/{\mu}l$, Myometrium tissue: $0.01531{\mu}g/{\mu}l$) and post-ovulatory stage(Epithelium tissue: $0.01414{\mu}g/{\mu}l$, Myometrium tissue: $0.01429{\mu}g/{\mu}l$). However, expression of u-PA did not differ from each estrous cycle in the epithelium tissue and myometrium tissue(p<0.05). Expression of HSP-90 was differ t-PA and u-PA from pre-ovulatory in Epithelium tissue($25,423{\mu}g/{\mu}l$) and early to mid-luteal stage in epithelium tissue($177,922{\mu}g/{\mu}l$) and myometrium tissue($26,664{\mu}g/{\mu}l$). These results suggest that HSP-90 and u-PA were related with change of uterus cycle according to the reformation of the tissues in porcine uterus.

Effects of pre-heat treatments on the quality of cut kimchi cabbages during short-term storage (절단배추의 단기 저장에 대한 예열처리 효과)

  • Kim, Sang-Seop;Ku, Kyung-Hyung;Jeong, Moon-Cheol;Hong, Joo-Heon;Chung, Shin-Kyo
    • Food Science and Preservation
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    • v.21 no.6
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    • pp.776-783
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    • 2014
  • Kimchi cabbages were cut ($3{\times}3cm$), and were pre-heat treated at $40^{\circ}C$, and their physicochemical qualities and browning degrees were investigated during 8 weeks storage at $5^{\circ}C$. The Cut kimchi cabbages were treated at $40^{\circ}C$ (1~8 hrs) and their protein bands profiles were determined by sodium dodecyl sulfate polyacrylamide electrophoresis (SDS-PAGE). The 60, 39, 33, and 12 kDa bands considered heat shock proteins (HSPs) were expressed in the cut kimchi cabbage, and the 4-hr pre-heat treatment (HS 4) exhibited the strongest band ratio. The weight ratios and titratable acidities of the pre-heat treated cut kimchi cabbages were not changed so much after 8 weeks storage at $5^{\circ}C$, and the soluble solid contents of HS 4 decreased less than that of any other treatments. The browning degree of HS 4 after 8 week storage was also shown to be the least among the treatments. The polyphenol oxidase (PPO) activities of all treatments slightly rose during the over all storage period, in contrast with the decrease of total phenolic contents. The expression of HSPs was identified in the pre-heat treated cut kimchi cabbages, and HS 4 exhibited the best quality and appearance after 8 weeks storage at $5^{\circ}C$.

Identification of Heat Stress-related Proteins and Low Molecular Weight HSP Expressed in Stem Tissues of Rice Plants by Proteomic Analysis (프로테옴 분석법에 의한 벼 줄기에서 발현하는 고온 스트레스 관련 단백질 및 저분자량 Heat Shock Protein의 분리 동정)

  • Lee, Dong-Gi;Kim, Kyung-Hee;Kim, Yong-Gu;Lee, Ki-Won;Lee, Sang-Hoon;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.31 no.2
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    • pp.99-106
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    • 2011
  • In order to investigate rice stem proteome in response to heat stress, rice plants were subjected to heat treatment at 42$^{\circ}C$ and total soluble proteins were extracted from stem tissues, and were fractionated with 15% PEG (poly ethylene glycol) and separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). After staining of 2-DE gels, 46 of differentially expressed proteins were extracted, digested by trypsin, and subjected to matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis. Proteins were identified through database search by using peptide mass fingerprints. Among them, 10 proteins were successfully identified. Seven proteins were up- and 3 proteins were down-regulated, respectively. These proteins are involved in energy and metabolism, redox homeostasis, and mitochondrial small heat shock proteins. 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, and also useful to molecular breeding of thermotolerant forage crops.

Proteome Analysis of Bacillus subtilis When Overproducing Secretory Protein

  • Jang Mi;Park Byoung-Chul;Lee Do-Hee;Kho Chang-Won;Cho Sa-Yeon;Lee Baek-Rak;Park Sung-Goo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.368-373
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    • 2006
  • Bacillus subtilis and related Bacillus species are frequently used as hosts for the mass production of recombinant proteins. Accordingly, this study examined the cellular response of B. subtilis to the overexpression of a soluble secretory protein. As such, the lichenase derived from B. cereus was overexpressed in B. subtilis, initially localized in the cytoplasm as a mature form and then secreted into the medium. Thereafter, the proteome of B. subtilis was analyzed using 2D electrophoresis and MALDI-TOF mass spectrometry. The expression of several heat-shock proteins, such as dnaK and groEL, was increased under this condition. In addition, manganese superoxide dismutase and NADH dehydrogenase were also upregulated in the lichenase-secreting B. subtilis. Therefore, it was concluded that the transient accumulation of a secreted protein in B. subtilis before secretion acted as a stress on the cell, which in turn induced the expression of various protective proteins.