• Title/Summary/Keyword: 단백질 특성 분석

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한우체세포 복제란이식우 및 정상우의 분만전후 태반특성 및 단백질분리양상

  • 성환후;우제현;임석기;정학재;김봉기;최재혁;장유민;박수봉;윤종택
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.37-37
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    • 2003
  • 본 연구실에서는 체세포복제 수정란 이식우의 분만전후 태아와 모체 및 태반과의 내분비적인 관계를 검토하기 위해 복제수정란 이식우의 분만 전후에 있어서, 혈중progesterone 과 TGF-$\beta$$_1$ 수준을 분석한 결과 정상우의 분만직전과 달리 혈중progesterone 및 TGF-$\beta$$_1$ 농도는 감소되지 않고 높은 수준을 그대로 유지되어 복제수정란이식우의 경우 정상적인 분만 signal이 나타나지 않는 것으로 나타났다. 본 연구는 체세포 복제소 수정란 이식우에 있어서 태반조직의 특성을 정상우의 동일한 시기의 태반과 비교 검토하였으며 분만시기에 있어서 태반의 역할을 검토하였다. 태반은 복제란 이식우 및 정상우 임신우를 술을 실시하여 태반을 채취하여 실험에 공시하였다. 정상우의 임신기간중 혈중 progesterone 농도는 임신초기와 중기에 다소 증가되어 유지되다가 임신말기에는 급격한 progesterone의 감소현상과 더불어 분만이 유기되는 것으로 나타났으나 복제우의 경우 분만예정일 직전까지 progesterone의 급격한 감소는 일어나지 않고 높은 농도로 유지되는 현상을 보였다. 이때 제왕절제 수술중 제대근방에 있는 태반궁부를 적출하여 중량을 측정한 결과 정상우의 궁부에 비해 유의적으로 무거웠으며, 이것은 태아의 중량과 정의 상관관계가 있음이 확인되었으며 태아 및 궁부의 중량이 무거울수록 분만 전후의 태아사망율이 높은 것으로 나타났다 이때의 궁부를 조직학적으로 검토한 결과, 복제우의 태반궁부는 정상우의 궁부에 비해 cytotrophoblast 및 syncytiotrophoblast가 적었으며 치밀하지 못한 형태를 확인할 수 있었다. 이때의 태반조직를 이용하여 세포질을 추출하여 단백질양상을 검토한 결과, 정상태반에 비해 복제태반은분자량 90kd, 65kd 및 18kd의 특이단백질이 존재하고 있음을 확인할 수 있었으며 이들 단백질을 HPLC system에 의해 분리한 결과 fraction 2와 3에서 특이한 단백질을 분리하였다. 이와 같이, 복제우의 태반에는 정상적인 임신과 분만을 비롯하여 복제송아지의 생존에 중요한 인자가 관여하고 있을 확인할 수 있었다.

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Cloning and Characterization of dnaK Heat Shock Protein Gene in a Halotolerant Cyanobacterium (내염성 cyanobacteria로 부터 danK heat shock protein 유전자의 cloning 및 특성 해명)

  • ;;;Teruhiro Takabe
    • Journal of Life Science
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    • v.11 no.5
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    • pp.464-469
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    • 2001
  • A gene, dnaK2, encoding a distinct member of the HSP70 family of molecular chaperones is isolated from the halotolerant cyanobactrium Aphanothece halophytica. The dnak2 gene encodes a molecular wight of 68 kDa polypeptide with predicted 616 amino acid residues. The DnaK2 protein has a structural characteristic of bacterial DnaK homologues and shows high similarity to other HSP70/Dank proteins. The danK2 transcripts are hardly detectable at 28$^{\circ}C$ and strongly induced upon heat stress. It is also found that dnaK2 transcript is increased by high-salinity stress even in the absence of heat stress. These results suggest that the DnaK2 protein plays an important role in protecting A. halophytica against damage caused by salt stress at well as heat stress.

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Assessing Grain Quality Properties of Reclaimed Wastewater Irrigation (하수처리수의 농업용수 재이용에 따른 벼의 미질특성 분석)

  • Jung, Ki-Woong;Kim, Hak-Kwan;Jang, Tae-Il;Kim, Sang-Min;Park, Seung-Woo
    • Journal of agriculture & life science
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    • v.44 no.6
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    • pp.183-189
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    • 2010
  • This study assess the effects of reclaimed wastewater irrigation on grain quality-related properties of milled rice. To conduct the effects of wastewater reuse on rice quality, the contents of amylose, protein, and milled head rice recovery were analyzed. The water and the soil quality were monitored, and the grain characteristic was analyzed using a NIRS (Near Infra-Red Spectroscopy). The value of the grain properties of the rice which was cultivated with the reclaimed wastewater didn't show a big difference from other common rice. The significant difference by each processing distinction was shown at the significance level of 5% in the statistical result.

Characterization of antimicrobial proteins produced by Bacillus sp. N32 (Bacillus sp. N32 균주가 생산하는 항균 단백질 특성)

  • Lee, Mi-Hye;Park, In-Cheol;Yeo, Yun-Soo;Kim, Soo-Jin;Yoon, Sang-Hong;Lee, Suk-Chan;Chung, Tae-Young;Koo, Bon-Sung
    • The Korean Journal of Pesticide Science
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    • v.10 no.1
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    • pp.56-65
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    • 2006
  • An antagonistic bacterial isolate, that inhibits the growth of plant pathogens, was selected and identified from 5,000 isolates screened from the rhizosphere of various crop plants. An isolate Bacillus sp. N32, tested against Colletotrichum gloeosporioides causing anthracnose disease in hot pepper, produced both a heat resistant antifungal protein and a heat sensitive antifungal protein. The heat resistant protein was partially purified by Ammonium sulfate fractionation and gel filtration chromatography. The bioautography showed that the proteins possessed high antifungal activity. The biosynthetic gene cluster responsible for the heat resistant antifungal protein was cloned from cosmid library using DNA probe obtained from PCR product with the primers targeting the conserved nucleotide sequence of the synthetic genes reported earlier, Most of the clones obtained showed higher homology to fengycin antibiotic synthetic gene family reported earlier. On the other hand, the heat sensitive protein was isolated from SDS-PAGE and electroblotting to determine the N-terminal amino acid sequences. The heat sensitive antifungal protein gene was cloned from the ${\lambda}-ZAP$ libraries using a DNA probe based on the N-terminal amino acid sequences of the heat sensitive protein. We are contemplating to clone and sequence the whole gene cluster encoding the heat sensitive protein for further analysis.

Protein-Protein Interaction Reliability Enhancement System based on Feature Selection and Classification Technique (특징 추출과 분석 기법에 기반한 단백질 상호작용 데이터 신뢰도 향상 시스템)

  • Lee, Min-Su;Park, Seung-Soo;Lee, Sang-Ho;Yong, Hwan-Seung;Kang, Sung-Hee
    • The KIPS Transactions:PartB
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    • v.13B no.7 s.110
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    • pp.679-688
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    • 2006
  • Protein-protein interaction data obtained from high-throughput experiments includes high false positives. In this paper, we introduce a new protein-protein interaction reliability verification system. The proposed system integrates various biological features related with protein-protein interactions, and then selects the most relevant and informative features among them using a feature selection method. To assess the reliability of each protein-protein interaction data, the system construct a classifier that can distinguish true interacting protein pairs from noisy protein-protein interaction data based on the selected biological evidences using a classification technique. Since the performance of feature selection methods and classification techniques depends heavily upon characteristics of data, we performed rigorous comparative analysis of various feature selection methods and classification techniques to obtain optimal performance of our system. Experimental results show that the combination of feature selection method and classification algorithms provide very powerful tools in distinguishing true interacting protein pairs from noisy protein-protein interaction dataset. Also, we investigated the effects on performances of feature selection methods and classification techniques in the proposed protein interaction verification system.

Oxalate Chelating Activity of Egg White Proteins and Their Hydrolysates

  • Holipitiyage Shyami Rashmiki, Holipitiya;Palihawadanege Iresha Lakmini, Fernando;Ethige Chathura Nishshanka, Rathnapala;Alakolange Gedara Achala Wimukthika, Alakolanga;Edirisinghe Dewage Nalaka Sandun, Abeyrathne;Ki-Chang, Nam
    • Korean Journal of Poultry Science
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    • v.49 no.4
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    • pp.221-228
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    • 2022
  • Major egg white proteins and their hydrolysates serve as functional food ingredients that have certain metal-chelating properties. Employing egg white proteins and their hydrolysates to scavenge dietary oxalates is anticipated to have beneficial effect in the prevention of kidney stones. The objective of this study was to determine the biogenic oxalate-chelating activity of ovalbumin, ovomucin, and ovotransferrin and their hydrolysates. To prepare oxalate extracts, 30 mL of 0.25 N HCl was added to separately to 0.5 g of dried spinach and starfruit powders followed by boiling for 15 min, and after cooling, the addition of a further 20 mL of 0.25 N HCl. Having prepared these extracts, ovalbumin, ovomucin, and ovotransferrin and their hydrolysates were separately mixed with oxalate extracts and incubated at 3℃ for 24 h. Following centrifugation, supernatants were analyzed by HPLC using a reverse-phase C18 column coupled with a diode array detector. We found that all assessed proteins and their hydrolysates showed biogenic oxalate-chelating activity against the oxalates of spinach. In contrast, however, only ovalbumin, ovalbumin-hydrolysate, and ovomucin showed chelating activity (57.10%±8.84%, 85.44%±5.30%, 73.20%±4.13%, respectively) against the oxalates of starfruit (P<0.05). Overall, hydrolyzed ovalbumin was identified as the most effective chelator of the oxalates both spinach and starfruit. In this study, we thus established that the assessed egg white proteins and their hydrolysates have oxalate-chelating activity in vitro, thereby indicating that these compounds have potential utility as nutraceuticals for the chelation of dietary oxalate. However, further research will be necessary to verify their oxalate-chelating activities against different fruits and vegetables and under specific in vivo conditions and against purified oxalate.

Nucleotide Sequence of an Extracellular Phospholipase D Gene from Streptomyces somaliensis and Transphosphatidylation Activity of Its Enzyme (Streptomyces somaliensis가 생산하는 세포외 Phospholipase D의 유전자 서열 분석과 Transphosphatidylation 활성 특성)

  • Jeong Sujin;Lee Sun-Hee;Uhm Tai-Boong
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.211-216
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    • 2004
  • A bacterial strain JE-ll found to produce active extracellular phospholipase D (PLD) was selected from the soil isolates. It was identified as Streptomyces somaliensis on the basis of 16S rDNA sequence analysis, morphological and physiological characteristics. The gene (sspld) encoding S. somaliensis PLD was isolated and characterized. The open reading frame was suggested to encode 538 amino acids with a signal peptide of 33 amino acids. The deduced amino acid sequence of the sspld shared a sequence similarity of 70-88% with PLDs of other Streptomyces sp. so far reported. The PLD converted phosphatidylcholine to phosphatidylglycerol or phosphatidylserine with the yield of 96 to 99% (㏖/㏖), but did not act on inositol or ethanolamine as a transphosphatidylation donor.

Molecular Analysis and Enzymatic Characterization of Cathepsin B from Olive Flounder (Paralichthys olivaceus) (넙치 카텝신 B의 분자생물학적 분석 및 효소학적 특성 연구)

  • Jo, Hee-Sung;Kim, Na-Young;Lee, Hyung-Ho;Chung, Joon-Ki
    • Journal of Fisheries and Marine Sciences Education
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    • v.26 no.3
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    • pp.543-552
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    • 2014
  • Papain family중 하나인 cysteine protease는 근골격계 질환 치료를 위한target molecule로 인식 되어왔으며 Cathepsin B 는 단백질 분해의 초기과정에 관여하는 cysteine proteases 중 하나이다. 본 연구는 넙치의 cathepsin B 유전자의 발현 양상과 넙치 cathepsin B(PoCtB)의 클로닝, 발현 및 효소특성을 분석하였다. cDNA Library Screening을 이용하여 넙치의 cDNA를 클로닝하였다. 넙치의 동정된 cathepsin B 유전자는 993bp의 open reading frame과 330개의 아미노산으로 이루어져있다. Cathepsin B의 propeptide region 내에 GNFD motif와 occluding loop 가 존재함으로써 이것이 명백하게 cathepsin B group이라는 것을 보여주며, 계통 유전학적 분석 결과 다른 종의 cathepsin B에 비해 초창기에 분화되어 나온 것으로 사료된다. mature enzyme인 maPoCtB은 fusion protein인 glutathione S-transferase를 포함하는 pGEX-4T-1 vector에 삽입하여 E.coli 균주인 $DH5{\alpha}$ 내에 발현시켰다. 재조합 단백질인 PoCtB을 과발현 시킨 결과 53kDa의 분자량을 가진다. 넙치 cathepsin B 활성은 Z-Arg-Arg-AMC와 같은 fluorogenic 펩타이드 기질을 이용하여 측정되었고 적정 pH는 pH.7.5 이다.

Comparison of Properties Affecting the Palatability of 33 Commercial Brands of Rice (시판 브랜드 쌀 33종의 품종별 식미 관련 특성 비교)

  • Kim, Chae-Eun;Kang, Mi-Young;Kim, Mi-Hyun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.3
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    • pp.301-309
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    • 2012
  • This study was performed to compare the properties affecting the palatability of 33 commercial brands of rice. Five rice varieties were identified including Chucheongbyeo, Hitomebore, Ilpumbyeo, Nampyeongbyeo, Seachucheongbyeo, were compared in terms of physicochemical characteristics, texture, head riceratio, and palatability through Toyo values. We also analyzed the relationship between grain characteristics and palatability. Amylose content of 5 rice varieties ranged from 17.04-17.98%. Nampyeongbyeo had the lowest and Seachucheongbyeo had the highest content of amylose among rice varieties. The protein content of 5 varieties ranged from 6.72-7.55%. Nampyeongbyeo showed the highest content. The moisture content varied from 13.08-14.83%. Chucheongbyeo has the highest moisture content. The head rice ratio of Ilpumbyeo, Chucheongbyeo, and Seachucheongbyeo were over the 90%, which was lower than that of Japan. Chucheongbyeo had the lowest hardness value and the highest adhesiveness value as measured by texture analyzer. The palatability value was highest in Chucheongbyeo, and the lowest in both Nampyeongbyeo and Ilpumbyeo. The palatability value was negatively correlated with protein content, but positively correlated with texture, moisture content, and head rice ratio. Based on this results, the rice varieties with high palatability had lower contents of amylose and protein, but higher moisture content and head rice ratio. Chucheongbyeo seemed to be the valuable variety with the highest palatability among them.

Characterization of Grain Amino Acid Composition and Proteome Profile of a High-lysine Barley Mutant Line M98 (고-Lysine 보리 돌연변이 계통 M98 종실의 아미노산 조성 및 Proteome Profile 특성)

  • Kim, Dea-Wook;Kim, Hong-Sik;Park, Hyoung-Ho;Hwang, Jong-Jin;Kim, Sun-Lim;Lee, Jae-Eun;Jung, Gun-Ho;Hwang, Tae-Young;Kim, Jung-Tae;Kim, Si-Ju;Rakwal, Randeep;Kwon, Young-Up
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.2
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    • pp.171-181
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    • 2012
  • Lysine is the first limiting essential amino acid in cereals for humans and monogastric animals, although its content is generally low. A chemically induced high-lysine barley mutant, M98, has an agronomically undesirable shrunken endosperm trait. In order to obtain detailed insight into the atypical traits of M98 grains, we characterized amino acid composition and protein profiles of M98 and its parent cultivar Chalssalbori. Among a total of 16 amino acids, the percentage of each of the 7 amino acids, including lysine, was 1.2~1.8 times higher in M98, comparing to Chalssalbori. The percentage of proline and its precursor, glutamic acid, in M98 was about the half of that of the amino acids in Chalssalbori, but arginine synthesized from glutamic acid was 1.8 times higher in M98, compared that in the parent cultivar. Theses results indicated that the mutation in M98 grains might alter the proportion of amino acids linked to each other in a biosynthetic pathway. A comparison of grain proteome profiles between Chalssalbori and M98 revealed 70 differentially expressed protein spots, where 45 protein spots were up-regulated and 25 protein spots down-regulated in M98 compared to those in Chalssalbori. Of these changed protein spots, 53 were identified using nano-electrospray ionization liquid chromatography mass spectrometry. Most of these identified proteins were involved in various biological processes. In particular, 28 protein spots such as ${\beta}$-amylase, serpins and B3-hordein were identified as proteins associated with the atypical traits of M98. It was thought that a genetic study on the unique protein profile of M98 would be needed to develop an agronomically feasible barley cultivar with high-lysine trait.