• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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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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• 제38권6호
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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.

• 원예과학기술지
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• 제30권5호
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• pp.509-518
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• 2012

이번 연구는 새싹채소 재배 시 고온의 물을 이용한 열처리가 케일 새싹의 항산화물질과 항암성물질 등과 같은 기능성물질 축적에 영향을 주는지 확인하고자 수행되었다. 실험I 에서는 파종 후 4일째 40, 50, $60^{\circ}C$의 증류수가 담긴 항온조에 각각 10, 30, 60초 동안 침지하여 열처리 하였고, 실험II 에서는 $50^{\circ}C$의 증류수에 각각 10, 20, 30, 45, 60초 침지처리 하였다. 열처리 한 케일 새싹은 다시 정상적인 생육 환경에서 2일 동안 추가적으로 재배되었다. 열처리 전, 후에 생체 중, 건물중, 전해질 유출 정도, 총 페놀 농도, 항산화도, 총 플라보노이드 농도, 페닐알라닌 암모니아-리아제 활성, 글루코시놀레이트 함량을 측정하였다. 실험I의 결과, 열처리 후 2일째 대조구에 비해 모든 처리구의 생체중, 건물중이 감소되었으며, 특히 $60^{\circ}C$ 열처리가 40, $50^{\circ}C$의 열처리보다 그 정도가 심했다. 세포 파괴에 의한 전해질 유출 정도 또한 $60^{\circ}C$에서 가장 높게 나타났다. 총 페놀 농도는 $50^{\circ}C$의 열처리에서 대조구에 비해 유의적으로 증가했으며 항산화도 또한 비슷한 경향을 보였다. 실험 II의 결과 $50^{\circ}C$ 열처리는 실험I의 결과와 같이 생체중의 감소를 보였지만 그 정도는 크지 않았으며 건물중에서는 대조구와 유의적 차이가 없었다. 열처리 후 2일째 모든 처리구는 대조구에 비해 유의적으로 높은 총 페놀 축적을 유도하였다. $50^{\circ}C$에서 20초 이상의 열처리는 대조구에 비해 약 1.5배 이상 총 페놀 농도를 증가시켰으며, 항산화도는 대조구에 비해 약 1.2배 유의적으로 높았고, 총 플라보노이드 농도 또한 대조구에 비해 높은 수준을 나타냈다. 페닐알라닌 암모니아-리아제의 활성은 열처리 24시간째에 대조구보다 모든 처리구에서 높게 나타나, 열처리가 이차대사산물의 생합성을 유도하는 것을 알 수 있었다. 열처리 후 2일째 글루코시놀레이트의 함량은 20초 열처리에서 유의적으로 가장 높은 값을 나타냈다. 따라서, 케일 새싹 재배 시 열처리는 항산화 및 항암물질과 같은 기능성물질의 축적을 유도하여 케일 새싹의 영양학적 품질향상을 도모할 수 있는 잠재적인 수단임을 확인할 수 있었다.

• 한국패류학회지
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• 제29권3호
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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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• 제7권1호
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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.

• 생명과학회지
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• 제19권12호
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• pp.1705-1711
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• 2009

항암제 내성을 획득한 암세포는 많은 경우 다양한 세포 독성 물질에 대해 교차 내성을 나타낸다. 그러나 온열 치료가 내성 획득 종양에 적용 될 때의 종양 세포의 사멸 효과는 알려져 있지 않다. 본 연구는 시스플라틴에 내성을 갖는 위암 세포 주, SNU601/Cis2이 열충격에 반응하는 민감도와 세포 사멸 방식을 조사함으로써 약물 내성 종양의 온열 치료 효과를 예측하고자 하였다. 정상 위암 세포 주 SNU601/WT은 열충격에 매우 민감하게 반응하며 apoptosis로 사멸하지만, 내성 위암 세포 주 SNU601/Cis2는 미열충격에 내성을 나타내었으며 고열충격에 노출되자 necrosis로 사멸하였다. 또한 SNU601/Cis2에서 necrosis의 발생은 열충격에 의한 JNK1/2의 활성화와 HSP27의 발현저하 현상과 관련되어 있었다. Necrosis의 유도는 세포막 파괴에 의해 세포 내부 물질의 방출로 인한 주변 조직의 염증반응을 수반하는데, 이러한 염증 반응은 암의 성장을 촉진하고 암의 성상을 심화시키는 것으로 보고되고 있다. 이러한 관점에서, 온열 치료가 약물 치료와 병행 될 경우에는 교차 내성과 necrosis로 인한 역효과를 방지하기 위하여, 그 적용이 주의 깊게 이루어져야 할 것으로 판단된다.

• 대한약리학회:학술대회논문집
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• 대한약리학회 2006년도 58th Annual Meeting of The Korean Society of Pharmacology
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• pp.42-42
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• 2006

• 한국식물학회:학술대회논문집
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• 한국식물학회 1993년도 제7회 식물생명공학 심포지움 식물 세포 분화의 분자적 접근 Seventh Symposium on Plant Biotechnology -Approach to Plant Cell Differentiation-
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• pp.113-148
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• 1993

• 한국수정란이식학회지
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• 제16권1호
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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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• 제7권1호
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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.