• 생명과학회지
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• 제20권12호
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• pp.1743-1749
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• 2010

Edwardsiella tarda는 그람 음성의 장내세균과의 주요 어병세균으로 어류에 edwardsiellosis를 유발하는 전신감염성 병원체이다. 최근 병원성 세균의 외막 단백질들은 세균성 감염에 있어서 숙주와 반응하여 면역반응을 유도하는 것으로 여겨져 연구가 되고 있다. 일본의 연구팀은 어류에서 에드워드병의 원인체인 E. tarda의 37 kDa 단백질이 넙치에서 높은 항원성을 제시하는 것을 보고하였다. 또한 그 연구자들은 37 kDa 단백질의 N-말단 아미노산 서열이 GAPDH와 대응하는 것을 밝혔다. 본 연구에서는 다른 세균에서 알려진 N-말단 서열을 기반으로 primer를 제작하여 이에 상응하는 E. tarda DNA를 증폭하고 클로닝하였다. 이 DNA단편의 염기서열은 예상한 바와 같이 세균의 GAPDH유전자인 gapA와 높은 상동성이 있고, E. tarda GAPDH (etGAPDH)의 아미노산 서열은 다른 장내세균의 GAPDH와 70% 이상의 상동성을 보이는 것을 확인하였다. E. tarda의 외막단백질에 특이적으로 반응하는 항체를 이용하여 E. tarda의 GAPDH가 외막에 존재한다는 것을 증명하였고, gapA의 염기서열을 바탕으로하여 재조합 GAPDH를 과발현 시켰다. 과발현된 재조합단백질 GAPDH는 GAPDH 특이적인 항체를 제조하는데 사용되었고, 또한 넙치에 면역시켜 단일 단백질 백신으로서의 활용도를 모색하였다. 비록 재조합 GAPDH가 면역된 넙치에서 GAPDH에 특이적인 항체가 증가하였음에도 불구하고, E. tarda로 공격실험을 하였을 때 면역된 넙치의 생존율이 12.5%로 측정되어 면역된 그룹과 면역되지 않은 그룹간에 큰 차이가 없는 것이 확인되었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권3호
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• pp.248-253
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• 2005

AfsR2, originally identified from Streptomyces lividans, is a global regulatory protein which stimulates antibiotic biosynthesis. Through its stable chromosomal integration, the high level of gene expression of afsR2 significantly induced antibiotic production as well as the sporulation of S. lividans, implying the presence of yet-uncharacterized AfsR2-target proteins. To identify and evaluate the putative AfsR2-target proteins involved in antibiotic regulation, the proteomics-driven approach was applied to the wild-type S. lividans and the afsR2-integrated actinorhodin overproducing strain. The 20 gel-electrophoresis gave approximately 340 protein spots showing different protein expression patterns between these two S. lividans strains. Further MALDI-TOF analysis revealed several AfsR2-target proteins, including glyceraldehyde-3-phosphate dehydrogenase, putative phosphate transport system regulator, guanosine penta phosphate synthetase/polyribonucleotide nucleotidyltransferase, and superoxide dismutase, which suggests that the AfsR2 should be a pleiotropic regulatory protein which controls differential expressions of various kinds of genes in Streptomyces species.

• 한국가금학회지
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• 제33권4호
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• pp.249-254
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• 2006

본 연구는 재래닭의 성장 관련 유용 유전자를 검색하기 위하여, 재래닭에서 발현되는 유전자와 코니쉬에서 발현되는 유전자를 subtraction하여 cDNA library를 구축하고, 염기서열을 밝혀서 재래닭 특이 유용 유전자를 검증하고자 하였다. cDNA library에서 얻은 clone들의 염기서열을 분석하여 나타난 결과를 5개의 clone을 비교 분석하였다. Clone NDS-1(618nt)은 비록 타 종과의 상동성은 낮으나(10%) 해당 과정에서 중요한 역할을 담당하는 triosephosphate isomerase이며, Clone NDS-6(651nt)는 자에서 해당 과정에 관여하는 glyceraldehyde-3-phosphate dehydrogenase로 여겨진다. 그러나 3가지 유전자(clone NDS-2, NDS-10, NDS-12)는 다른 유전자들과 비교하여 5.0%내외의 낮은 상동성을 보이고 고등 동물과 거의 유사성이 없으므로 재래닭 특이 성장 관련 유용 유전자일 가능성이 있다.

• 한국식품저장유통학회지
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• 제15권1호
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• pp.84-87
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• 2008

유전자 변형 작물은 생산성 측면에서 많은 장점이 있지만 이를 섭취할 경우 잠재적인 위험 요소들에 의해 많은 문제가 대두대고 있다. 본 연구는 저항성유전자를 이입한 배추에서 Profillin, Tubulin-${\alpha}$ (Tub-${\alpha}1$), Heat-shock protein (Bchsp 17.6) and Ubiquitin conjugating enzyme (UBE)의 발현과 이를 30일간 섭취한 마우스에서 ${\beta}$-actin(${\beta}$-act), ${\beta}$-2-microglobulin (B2m), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ${\beta}$-glucuronidase (Gus)의 발현 정도를 RT-PCR을 통해 알아보았다. 실험 결과 저항성유전자를 이입한 배추와 그렇지 않은 배추의 유전자 발현 패턴은 큰 차이를 보이지 앓았으며, 이를 섭취한 마우스 장기에서도 발현에 따른 큰 차이는 나타나지 않았다.

• 한국균학회지
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• 제46권4호
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• pp.459-466
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• 2018

2017년 1월 정읍의 한 농가 저장창고에서 검게 변한 귀리종자가 발견되었다. 조사된 100개의 종자 중 45개 종자에서 Pyrenophora속 균이 검출되었다. 종자에서 얻어진 모든 균주들은 internal transcribed spacer (ITS) 부위와 glyceraldehyde 3-phosphate dehydrogenase (GPDH) 유전자 염기서열을 기초로 Pyrenophora avenae로 동정되었고, 형태적, 배양적 특성에 의해 확인되었다. ITS와 GPDH 염기서열 기반의 계통수를 통해 P. avenae 균주는 유전적으로 뚜렷한 4개의 그룹으로 구분할 수 있었다. 병원성 검정 결과, P. avenae는 귀리의 종자흑변과 잎마름병을 일으키는 병원균으로 확인되었다. 이것은 한국에서 P. avenae가 귀리잎마름병을 일으킨다는 최초 보고이다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권3호
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• pp.173-178
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• 2001

Food yeast, Saccharomyces cerevisiae, is a safe organism with a long history of use for the production of biomass rich in high quality proteins and vitamins. AmA1, a seed storage albumin from Amaranthus hypochondriacus, has a well-balanced amino acid composition and high levels of essential amino acids and offers the possibility of further improving food animal feed additives. In order to find an effective means of expressing AmA1 in yeast, the gene was cloned into an episomal shuttle vector. Four different promoters were tested: the glyceraldehyde-3-phosphate dehydrogenase promoter, galactose dehydrogenase 10 promoter, alcohol dehydrogenase II promoter, and a hybrid ADH2-GPD promoter. The recombinant AmA1 genes were then introduced into the yeast Saccharomyces cerevisiae 2805. Northern and Western blot analyses of the yeast under appropriate conditions revealed that AmA1 was expressed by all four promoters at varying levels. An enzyme-linked immunosorbent assay demonstrated that the amount of AmA1 protein in the recombinant yeast was 1.3-4.3% of the total soluble proteins. The highest expression level was obtained from the hybrid ADH2-GPD promoter.

• Korean Journal of Acupuncture
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• 제29권2호
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• pp.260-270
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• 2012

Objectives : Hippocampus, a region of temporal lobe, plays an important role in the pathogenic mechanisms of brain diseases such as Alzheimer's disease, depression and temporal lobe epilepsy. This research is designed to investigate hippocampal changes after acupuncture stimulation at Shinmun(HT7) using 2-dimensional gel electrophoresis(2-DE). Methods : On postnatal-day 15, rat pups were randomly devided into Normal(NOR) or HT7 group. All of Pups kept with their mothers for 7 days, but pups in HT7 group received acupuncture stimulation at HT7 daily. On postnatal-day 21, hippocampus of each rat pup was dissceted 30 minutes after last acupuncture stimulation and the protein expressions were investigated using 2-DE. Results : After acupuncture stimulation at HT7, expression of 20 proteins were significantly increased. Succinate semialdehyde dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase-like, transketolase, aconitate hydratase and phosphoglucomutase-1 were related to glucose methabolism. Eukaryotic initiation factor(eIF) 4A-II, eIF 4A-III, mitochondrial Tu translation elongation factor and chain A of crystal structure of the 70-Kda heat shock cognate protein involve in the protein synthesis in ribosome. Tubulin ${\beta}$-4 chain, tubulin T ${\beta}$-15 and tubulin ${\alpha}$-1B chain comprise cytoskeleton. Glutathione S-transferase(GST) ${\omega}$-1, GST P and GST Yb-3 can reduce oxidative stress. ${\beta}$-soluble N-ethylmaleimide-sensitive fusion protein attachment protein is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus, glycerol-3-phosphate dehydrogenase plays a major role in lipid biosynthesis, creatine kinase U-type catalyses the conversion of creatine and consumes adenosine triphosphate to create phosphocreatine and adenosine diphosphate. Platelet-activating factor acetylhydrolase IB subunit alpha and voltage depedent anion-selective channel protein 2 were also increased. Conclusions : The results suggest that acupuncture stimulation at HT7 may enhance glucose and lipid metabolism, protein synthesis, cytoskeletal substance and anti-oxidative stress in hippocampus.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권1호
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• pp.84-87
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• 2006

Batch binding experiments were performed to assess the recovery performance of glyceraldehyde 3-phosphate dehydrogenase (G3PDH) bound to the unshielded and polymer (polyvinyl pyrrolidone. PVP)-shielded dye-ligand (Cibacron Blue 3GA) adsorbent. The adoption of a polymer-shielded, dye-ligand technique facilitated the elution efficiency of bound G3PDH. It was demonstrated that the recovery of G3PDH using polymer-shielded dye-ligand adsorption yielded higher elution efficiency, at 60.5% and a specific activity of 42.3 IU/mg, after a low ionic strength elution (0.15 M NaCl). The unshielded dye-ligand yielded lower elution efficiency. at 6.5% and a specific activity of 10.2 IU/mg.

• 한국양식학회지
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• 제22권1호
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• pp.51-55
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• 2009

Transcriptional response of the second isoform of glyceraldehyde (GAPDH-2) to infectious challenges using various bacterial species and the rockbream iridovirus (RBIV) was examined in the spleen of the barred knifejaw (Oplegnathus fasciatus). Bacterial challenges of the juvenile barred knifejaws with Escherichia coli, Edwardsiella tarda, Vibrio anguillarum and Streptococcus iniae resulted in the significant elevation of the GAPDH-2 transcripts in the spleen up to 12-fold relative to that in the non-challenged controls (PBS-injected). In addition, the barred knifejaw fingerlings injected with RBIV stock also represented the highly upregulated mRNA expression of the GAPDH-2 up to more than 20-fold when compared to that of control fingerings. Results obtained from this study strongly suggest that the GAPDH-2 is no longer a housekeeping glycolytic protein and rather than that it might be associated with immune-relevant cellular activities. From this finding, the traditional dogma for the use of GAPDH as an invariant standard for gene expression assays should be carefully revised depending on the kinds of biological stimulations applied in this species.

• BMB Reports
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• 제44권12호
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• pp.782-786
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• 2011

Aberrant GAPDH expression following S-nitrosoglutathione (GSNO) treatment was compared in HepG2 cells, which express functional p53, and Hep3B cells, which lack functional p53. The results of Western blotting and fluorescent immunocytochemistry revealed that nuclear translocation and accumulation of GAPDH occur in both HepG2 and Hep3B cells. This finding suggests that p53 may not be necessary for the GSNO-induced translocation of GAPDH to the nucleus during apoptotic cell death in hepatoma cells.