• Molecules and Cells
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• 제23권1호
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• pp.100-107
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• 2007

The moss Physcomitrella patens has two life cycles, filamentous protonema and leafy gametophore. A modified from of suppression subtractive hybridization (SSH), mirror orientation selection (MOS), was applied to screen genes differentially expressed in the P. patens protonema. Using reverse Northern blot analysis, differentially expressed clones were identified. The identified genes were involved mainly in metal binding and detoxification. One of these genes was an AP2 (APETALA2) domain-containing protein (PpACP1), which was highly up-regulated in the protonema. Alignment with other AP2/EREBPs (Ethylene Responsive Element Binding Proteins) revealed significant sequence homology of the deduced amino acid sequence in the AP2/EREBP DNA binding domain. Northern analysis under various stress conditions showed that PpACP1 was induced by ethephon, cadmium, copper, ABA, IAA, and cold. In addition, it was highly expressed in suspension-cultured protonema. We suggest that PpACP1 is involved in responses to metals, and that suspension culture enhance the expression of genes responding to metals.

• 대한수의학회지
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• 제50권4호
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• pp.285-295
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• 2010

Protein expression patterns in Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium) strains with diverse phenotypes, such as phage type, antibiotic resistance pattern and plasmid profiles were examined. For detailed analysis of proteins expressed by different S. Typhimurium strains, protein fractions were divided into detergent-rich phase (DP) and aqueous phase (AP) using triton X-114 detergent. The two phases were subjected to two-dimensional gel electrophoresis (2-DE), followed by protein identification using peptide mass fingerprinting (PMF). In the results, PMF showed that DP fractions consisted mainly of outer membrane proteins, whereas the AP fractions included cytosolic proteins. Comparison of 2-DE profiles of DP did not show any distinct protein spots which could be correlated with phage type, antibiotic resistance pattern or plasmid profile. However, comparisons of 2-DE profiles of the AP revealed differences in the protein spots, which could be correlated with the plasmid profile and phage types. Among these protein spots, flagellin was specific for strains containing a 90 kb plasmid. Compared to DT193 phage type, three protein spots in the range of pI 5.0-5.5 and MW 8-15 kDa of AP 2-DE profiles were absent in the DT104 phage types. Additionally, a protein spot with PI in the range of 4.5-5.0 and molecular weight (MW) between 51-69 kDa was specific for phage type DT104, while a protein spot with pI in the range of 4.0-4.8 and MW between 18-20 kDa was specific for DT193 phage type. These protein spots may be useful for discriminating phage types of S. Typhimurium.

• 한국균학회지
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• 제26권4호통권87호
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• pp.502-510
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• 1998

장수버섯 자실체로부터 분리된 강력한 항보체 활성 다당체인 FF-AP1을 정제하고 이것의 화학적 특성을 구명하기 위해 본 실험을 수행한 결과는 다음과 같다. ConA-Sepharose 4B affinity chromatography에 의해 FF-AP1은 $FF-AP1{\alpha}$와 $FF-AP1{\beta}$로 더 분리되었으며 이 중 항보체 활성의 주 유효 다당체인 $FF-AP1{\beta}$는 $10\;{\mu}g/ml$ 농도에서도 약 70%의 항보체 활성도를 나타내었다. $FF-AP1{\beta}$는 약 9.6%의 단백질을 함유한 산성다당체이며 glutamic acid와 aspartic acid의 함량이 상대적으로 다른 아미노산들에 비해 높았다. 이것의 분자량은 gel filtration법에 의해 15,000인 것으로 추정되었고 적외선 분광 분석에서 $890cm^{-1}$ 부근의 소peak를 탐지함으로써 ${\beta}$ glucosidic 결합의 존재를 확인하였다. Gas chromatography에 의한 구성당 조성 분석 결과, 주 구성당이 glucose, mannose, 및 galactose인 galacto-manno-glucan이었고, 소량의 rhamnose, xylose, fucose와 uronic acid도 탐지되었다.

• Molecules and Cells
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• 제23권3호
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• pp.316-322
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• 2007

Zinc finger transcription factor genes are a significant fraction of the genes in the vertebrate genome. Here we report the isolation and characterization of a human zinc finger-containing gene, ZNF435, from a fetal brain cDNA library. ZNF435 cDNA is 1290 base pairs in length and contains an open reading frame encoding 349 amino acids with four C2H2-type zinc fingers at its carboxyl terminus and a SCAN motif at its amino terminus. RT-PCR results showed that ZNF435 was expressed in all tested tissues. A ZNF435-GFP fusion protein was located in the nucleus and the four zinc fingers acted as nuclear localization signals (NLSs). ZNF435 was found to be capable of homo-association, and this effect was independent of its zinc fingers. Furthermore, ZNF435 proved to be a transcription repressor as its overexpression in AD293 cells inhibited the transcriptional activities of AP-1.

• Molecules and Cells
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• 제28권6호
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• pp.545-551
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• 2009

Mitogen-activated protein kinases (MAPK) affect the activation of activator protein-1 (AP-1), which plays an important role in regulating a range of cellular processes. However, the roles of these signaling factors on hydrogen peroxide ($H_2O_2$)-induced cell death are unclear. This study examined the effects of $H_2O_2$ on the activation of MAPK and AP-1 by exposing the cells to $H_2O_2$ generated by either glucose oxidase or a bolus addition. Exposing BJAB or Jurkat cells to $H_2O_2$ affected the activities of MAPK differently according to the method of $H_2O_2$ exposure. $H_2O_2$ increased the AP-1-DNA binding activity in these cells, where continuously generated $H_2O_2$ led to an increase in mainly the c-Fos, FosB and c-Jun proteins. The c-Jun-$NH_2$-terminal kinase (JNK)-mediated activation of c-Jun was shown to be related to the $H_2O_2$-induced cell death. However, the suppression of $H_2O_2$-induced oxidative stress by either JNK inhibitor or c-Jun specific antisense transfection was temporary in the cells exposed to glucose oxidase but not to a bolus $H_2O_2$. This was associated with the disruption of death signaling according to the severe and prolonged depletion of reduced glutathione. Overall, these results suggest that $H_2O_2$ may decide differently the mode of cell death by affecting the intracellular redox state of thiol-containing antioxidants, and this depends more closely on the duration exposed to $H_2O_2$ than the concentration of this agent.

• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.59-64
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• 2002

An angiotensin I-converting enzyme (EC 3.4.15.1) (ACE), which can convert inactive angiotensin I into angiotensin II, a vasoconstrictor, is one of the key enzymes in controlling hypertension. It is suggested that the inhibition of ACE prevents hypertension, and many inhibitory peptides have already been reported. In the current study, oligonucleotides encoding ACE inhibitory peptides (IY, VKY) were chemically synthesized and designed to be multimerised due to isoschizomer sites (BamHI, BglII). The cloned gene named AP3 was multimerised up to 6 times in pBluescript and expressed in BL2l containing pGEX-KG. The fusion protein (GST-AP3) was easily purified with a high recovery by an affinity resin, yielding 38 mg of synthetic AP3 from a 1-1 culture. The digestion of AP3 by chymotrypsin exhibited an $IC_50$ value of $18.53{\mu}M$. In conclusion, the present experiment indicated that AP3 could be used as a dietary antihypertensive drug, since the potent ACE inhibitory activity of AP3 could be activated by chymotrypsin in human intestine.

• 생명과학회지
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• 제24권2호
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• pp.112-117
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• 2014

본 연구는 복분자와 쑥 추출물이 UVB (자외선-B)에 의한 유전독성의 감소에 효과를 보이는 지를 HaCaT 세포를 이용하여 분석하였다. 자외선을 조사하지 않은 정상 세포의 세포활성은 복분자와 쑥 추출물을 처리한 경우 처리하지 않은 대조군과 차이를 나타내지 않았지만, 자외선을 조사한 세포에 복분자와 쑥 추출물을 처리한 경우 처리하지 않은 대조군에 비해 농도 의존적으로 세포 활성을 증가하였다. UVB를 조사한 후 세포의 핵 분절율은 복분자와 쑥 추출물을 처리한 경우 정상 배양액으로 배양한 대조군에 비해 핵 분절율을 각각 약 20%, 15% 정도 감소하였다. DNA 상해 회복에 반응하는 cyclobutane pyrimidine dimer (CPD)의 잔여량을 확인한 결과, 복분자와 쑥 추출만 처리한 경우 처리하지 않은 대조군에 비해 차이를 나타내지 않았으나 UVB 조사한 후 복분자와 쑥추출물을 처리한 경우 정상 배양액으로 배양한 대조군에 비해 농도의존적으로 감소하였다. 세포 상해에 반응하는 유전자인 phospho-p53과 GADD45의 단백질 수준을 Western blot으로 분석한 결과, 자외선을 조사하지 않은 정상세포에 복분자와 쑥 추출물만 처리한 경우 처리하지 않은 대조군과 비교하여 유의적인 차이를 나타내지 않았지만 UVB 조사한 후 복분자와 쑥 추출물을 처리한 경우 농도 의존적으로 phospho-p53과 GADD45 단백질 발현량이 감소하였다. 위의 결과에서 복분자와 쑥 추출물은 UVB에 유도된 DNA 상해와 p53 및 GADD45와 같은 상해 반응단백질의 수준을 감소시키는 것으로 보아 UVB에 의해 손상된 세포의 유전독성을 회복하는 효능이 있다고 생각된다.

• BMB Reports
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• 제34권3호
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• pp.262-267
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• 2001

A cDNA coding alkaline phosphatase (AP) homologue was isolated from a cDNA library of Schizosaccharomyces pombe by colony hybridization. The nucleotide sequence of the cloned cDNA appeared to lack the N-terminal coding region. The genomic DNA encoding alkaline phosphatase homologue was isolated from S. pombe chromosomal DNA using PCR. The amplified DNA fragment was ligated into plasmid pRS315 to generate the recombinant plasmid pSW20. The DNA insert was subcloned as two smaller fragments for nucleotide sequencing. The sequence contains 2,789 by and encodes a protein of 532 amino acids with a molecular mass of 58,666 daltons. The S. pombe cells containing plasmid pSW20 showed much higher AP activity compared with the yeast cells with vector only This indicates that the cloned AP gene apparently encodes AP The predicted amino acid sequence of the S. pombe AP shares homology with those of other known APs.

• Journal of Microbiology
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• 제41권2호
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• pp.109-114
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• 2003

A promoter that is inducible by paraquat and menadione, the superoxide generators, independently of soxRS has been found in front of the sufABCDSE operon in Escherichia coli. Based on the observation that SufA is a holomog of IscA that functions in the assembly of iron sulfur cluster and the sufA promoter (sufAp) contains a putative Fur-binding consensus, we investigated whether this gene is regulated by Fur, a ferric uptake regulator, When examined in several sufAp-lacZ chromosomal fusion strains, sufAp was induced by EDTA, an iron chelator and a well-known Fur-inducer, The basal level of sufA expression increased dramatically in fur mutant, suggesting repression of sufAp by Fur. The derepression in fur mutant and EDTA-induction of sufA expression required nucleotides up to -61, where a putative Fur box is located. Purified Fur protein bound to the DNA fragment containing the putative Fur box between -35 and -10 promoter elements. The regulation by Fur and menadione induction of sufAp acted independently. The rpoS mutation increased sufA induction by menadione, suggesting that the stationary sigma factor RpoS acts negatively on sufA induction.

• 한국유기농업학회지
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• 제5권2호
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• pp.117-128
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• 1997

This study was carried out to investigate the amounts of voluntary intake, digestibility and nitrogen retention in Korean native goats (KNG) fed agricultural by-products containing rice straw (RS) and apple pomace (AP) and to obtain a basic information for establishing the feeding system of KNG. The result are as follows. 1. Among the chemical composition of experimental diets, the highest values in crude protein (CP ; 18.6%) and crude ash contents (10.7%) were observed in alfalfa hay. Those of RS+AP treatment were significantly low 6.0 and 4.8%, respecitively. Acid detergent fiber (ADF), ether extract and non-structural carbohydrate (NSC) contents were shown an adverse tendency. 2. Dry matter (DM) intakes per day in KNG fed RS+commercial diet (CD), RS+AP and RS+AP+CD were significantly lower (P<0.05) 210.3, 228.3, respectively than 358.1g in alfalfa hay. 3. DM intakes per basal weight expressed as DM g/kg of BW0.75 and DM g/kg of BW(%) were highest (P<0.05) 60.5g and 3.3%, respectively in KNG fed alfalfa hay, any other treatments showed 35.6 to 42.5g and 2.0 to 2.3%, respectively and this result was similar to those of DM intake per day. 4. Digestibilities of DM, organic matter, CP, ADF, neutral detergent fiber (NDF). crude ash and ether extract in alfalfa hay and RS+AP+CD treatment were significantly higher (P<0.05) than RS+CD treatment. Those of CP, ADF and NDF of RS+AP treatment in not significantly different with these treatments. 5. NSC digestibility was higher in alfalfa hay (66.7%) than those of other treatments (44.2~52.0% ; P<0.05). 6. Nitrogen retentions expressed as N retention(g) and N retention (%) in KNG were highest (P<0.05) 0.6g and 15.6%, respectively in RS+AP+CD treatment and RS+CD treatment was lowest (P<0.05) -06g and -21.4%, respectively.