• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.355-356
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• 2000

Three 'stress probe' plasmids were constructed and characterized which utilize a green fluorescent protein (CFP) as a non-invasive reporter to elucidate Escherichia coli cellular stress responses in quiescent or 'resting' cells. Facile detection of cellular stress levels was achieved by fusion of three heat shock stress protein promoter elements, those of the heat shock transcription factor ${\sigma}^{32}$, pretense subunit ClpB, and chaperone DnaK, to the reporter gene $gfp_{uv}$. When perturbed by chemical or physical stress (such as heat shock, nutrient (amino acid) limitation, addition of IPTG, acetic acid, ethanol, phenol, antifoam, and salt (osmotic shock), the E. coli cells produced GFPuv which was easily detected from within the cells as emitted green fluorescence. A temporal and amplitudinal mapping of these responses was performed, demonstrating regions where quantitative delineation of cell stress was afforded.

• BMB Reports
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• v.38 no.6
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• pp.695-702
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• 2005

The groESL bicistronic operon of a restricted facultative methylotrophic bacterium Methylovorus sp. strain SS1 DSM 11726 was cloned and characterized. It was found to consist of two ORFs encoding proteins with molecular masses of 11,395 and 57,396 daltons, which showed a high degree of homology to other bacterial GroES and GroEL proteins. The genes were clustered in the transcription order groES-groEL. Northern blot analyses suggested that the groESL operon is transcribed as a bicistronic 2.2-kb mRNA, the steady-state level of which was markedly increased by temperature elevation. Primer extension analysis demonstrated one potential transcription start site preceding the groESL operon, which is located 100bp upstream of the groES start codon. The transcription start site was preceded by a putative promoter region highly homologous to the consensus sequences of Escherichia coli ${\sigma}^{32}$-type heat shock promoter, which functioned under both normal and heat shock conditions in E. coli. Heat shock mRNA was maximally produced by Methylovorus sp. strain SS1 approximately 10min after increasing the temperature from 30 to $42^{\circ}C$. The groESL operon was also induced by hydrogen peroxide or salt shock.

• Horticultural Science & Technology
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• v.30 no.5
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• pp.509-518
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• 2012

The objective of this study was to determine the effect of heat shock treatments on the phytochemicals including antioxidants and anticancer materials in kale (Brassica oleracea L. var. acephala) sprouts. In study I, kale sprouts grown under the growing system for four days were soaked at 40, 50, or $60^{\circ}C$ distilled water for 10, 30, or 60 seconds, and in study II, kale sprouts were soaked at $50^{\circ}C$ distilled water for 10, 20, 30, 45, or 60 seconds. After the heat shock treatments, the sprouts were transferred into normal growing conditions and recovered there for two days. Fresh and dry weights, electrolyte leakage, total phenolic concentration, antioxidant capacity, total flavonoid concentration, phenylalanine ammonia-lyase (PAL) activity, and glucosinolates content of the sprouts were measured before and after the heat shock treatments. As a result, there was a significant decrease in the fresh and dry weight of kale sprouts treated with heat shock compared with control at harvest in study I. Especially, heat shock at $60^{\circ}C$ lead to more pronounced growth inhibition compared with heat treatments at 40 and $50^{\circ}C$. Electrolyte leakage by cell collapse was the highest in the sprouts exposed to $60^{\circ}C$ distilled water, which agreed with the growth results. Heat shock at $50^{\circ}C$ significantly induced the accumulation of phenolic compounds. In study II, fresh weight of kale sprouts at $50^{\circ}C$ heat shock showed a significant decrease compared with the control at one and two days after the treatment. However, the decrease was minimal and dry weight of kale sprouts was not significantly different from that in control. In contrast, the heat shock-treated kale sprouts had higher level of total phenolic concentration than control at harvest. Heat shock treatments at $50^{\circ}C$ for 20 seconds or more showed at least 1.5 and 1.2 times higher total phenolic concentration and antioxidants capacity than control, respectively. The change of the total flavonoid concentration was similar with that of antioxidants. PAL activity after 24 hours of heat shock was higher in all the heat shock-treated sprouts than that in control suggesting heat shock may stimulate secondary metabolic pathway in kale sprouts. Seven glucosinolates were identified in kale sprouts and soaking the sprouts with $50^{\circ}C$ water for 20 seconds had a pronounced impact on the accumulation of total glucosinolates as well as two major glucosinolates, progoitrin and sinigrin, at harvest. In conclusion, this study suggests that heat shock using hot water would be a potential strategy to improve nutritional quality of kale sprouts by inducing the accumulation of phytochemicals with antioxidant and anticancer properties.

• The Korean Journal of Malacology
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• v.29 no.3
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• pp.181-187
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• 2013

In order to investigate environmental stress inducible genes in abalone, we analyzed differentially expressed transcripts from a Pacific abalone, Haliotis discus hannai, after exposure to heat-, cold- or hyposalinity-shock by suppression subtractive hybridization (SSH) method. 1,074 unique sequences from SSH libraries were composed to 115 clusters and 986 singletons, the overall redundancy of the library was 16.3%. From the BLAST search, of the 1,316 ESTs, 998 ESTs (75.8%) were identified as known genes, but 318 clones (24.2%) did not match to any previously described genes. From the comparison results of ESTs pattern of three SSH cDNA libraries, the most abundant EST was different in each SSH library: small heat shock protein p26 (sHSP26) in heat-shock, trypsinogen 2 in cold-shock, and actin in hyposalinity SSH cDNA library. Based on sequence similarities, several response-to-stress genes such as heat shock proteins (HSPs) were identified commonly from the abalone SSH libraries. HSP70 gene was induced by environmental stress regardless of temperature-shock or salinity-stress, while the increase of sHSP26 mRNA expression was not detected in cold-shock but in heat-shock condition. These results suggest that the suppression subtractive hybridization method is an efficient way to isolate differentially expressed gene from the invertebrate environmental stress-response transcriptome.

• Mass Spectrometry Letters
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• v.7 no.1
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• pp.1-11
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• 2016

Various efforts have been developed to improve sample preparation steps, which strongly depend on hands-on processes for accurate and sensitive quantitative proteome analysis. In this study, we carried out heating the sample prior to trypsin digestion using an instrument to improve the tryptic digestion process. The heat shock generated by the system efficiently denatured proteins in the sample and increased the reproducibility in quantitative proteomics based on peptide abundance measurements. To demonstrate the effectiveness of the protocol, three cell lines (A human lung cancer cell line (A549), a human embryonic kidney cell line (HEK293T), and a human colorectal cancer cell line (HCT-116)) were selected and the effect of heat shock was compared to that of normal tryptic digestion processes. The tryptic digests were desalted and analysed by LC-MS/MS, the results showed 57 and 36% increase in the number of identified unique peptides and proteins, respectively, than conventional digestion. Heat shock treated samples showed higher numbers of shorter peptides and peptides with low inter-sample variation among triplicate runs. Quantitative LC-MS/MS analysis of heat shock treated sample yielded peptides with smaller relative error percentage for the triplicate run when the peak areas were compared. Exposure of heat-shock to proteomic samples prior to proteolysis in conventional digestion process can increase the digestion efficiency of trypsin resulting in production of increased number of peptides eventually leading to higher proteome coverage.

• Journal of Life Science
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• v.19 no.12
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• pp.1705-1711
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• 2009

Carcinoma cells that had acquired resistance to a chemotherapeutic drug often show cross-resistance to various other cytotoxic drugs. In the present study, we explored the effect of heat shock in cisplatin-resistant gastric cancer cells SNU601/Cis2 to figure out the efficacy of hyperthermia in drug-resistant carcinoma. While SNU601/WT cells showed a high-sensitivity response to heat shock by dying through apoptosis, SNU601/Cis2 cells were considerably resistant to mild heat shock, but died by necrosis upon treatment with harsh heat shock. The occurrence of necrosis in SNU601/Cis2 cells was linked to the suppression of both JNK1/2 activation and HSP27 induction in response to heat shock. Since necrosis is closely associated with tumor malignancy and poor prognosis through inflammatory responses, our result suggests that hyperthermic treatment should be carefully applied when it is combined with chemotherapy.

• 대한약리학회:학술대회논문집
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• 2006.11a
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• pp.42-42
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• 2006

• Proceedings of the Botanical Society of Korea Conference
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• 1993.07a
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• pp.113-148
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• 1993

• Journal of Embryo Transfer
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• v.16 no.1
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• pp.41-46
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• 2001

This study was carried out to evaluate the effect of culture methods on development of embryos with each developmental stage after heat shock in bovine oocytes. The results obtained were as fellows. 1. The culture method after heat shock on development of embryos was better drop-culture than co-culture. 2. The medium without amino acids were not effect of heat sock on development of embryos but it was in need of amino acid during formation of blastocyst.

• Genomics & Informatics
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• v.7 no.1
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• pp.26-31
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• 2009

Heat shock proteins are a class of molecular chaperones that can be found in nearly all organisms from Bacteria, Archaea and Eukarya domains. Heat shock proteins experience increased transcription during periods of heat induced osmotic stress and are involved in protein disaggregation and refolding as part of a cell's danger signaling cascade. Heat shock protein, Hsp20 is a small molecular chaperone that is approximately 20kDa in weight and is hypothesized to prevent aggregation and denaturation. Hsp20 can be found in several strains of Proteobacteria, which comprises the largest phyla of the Bacteria domain and also contains several medically significant bacterial strains. Genomic analyses were performed to determine a common evolutionary pattern among Hsp20 sequences in Proteobacteria. It was found that Hsp20 shared a common ancestor within and among the five subclasses of Proteobacteria. This is readily apparent from the amount of sequence similarities within and between Hsp20 protein sequences as well as phylogenetic analysis of sequences from proteobacterial and non-proteobacterial species.