• Title/Summary/Keyword: 단백질 동정

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The Comparison between FSGS and MCNS Using Proteomic Method in Childhood Nephrotic Syndrome; Preliminary Study (단백질체학을 이용하여 국소성 분절성 사구체 경화증과 미세 변화형 신증후군의 비교)

  • Kim, Sung-Do;Cho, Byoung-Soo
    • Childhood Kidney Diseases
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    • v.13 no.2
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    • pp.170-175
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    • 2009
  • Purpose : FSGS do not respond well to any kind of therapy and gradually progress to end-stage renal disease. This study was conducted to investigate the difference of protein expression between MCNS and FSGS as a preliminary study for understanding the pathophysiology of FSGS. Methods : Renal biopsy samples of MCNS and FSGS were obtained, which was diagnosed by one pathologist. They were solubilized with a conventional extraction buffer for protein extraction. The solution was applied on immobilized linear gradient strip gel (pH 4-7) using IPGphor system. Silver staining was carried out according to standard method. Protein identification was done by searching NCBI database using MASCOT Peptide Mass Fingerprint software. Results : The differences in protein expressions between MCNS and FSGS were shown by increased or decreased protein spots. Most prominently expressed spot among several spots in FSGS was isolated and analyzed, one of which was glutathione S-transferase (GST) P1-1, whereas it was not found in MCNS. So GSTP1-1 was considered as the one of the key biomarkers in pathogenesis of FSGS. Conclusion : This result would be helpful in diagnosing FSGS and researching FSGS. Further studies for glutathione S-transferase P1-1 might be necessary to elucidate the mechanisms regarding FSGS.

Transcriptional regulation of soybean ${\beta}-conglycinin$ gene expression. -(I) Identification of a soybean embryo factor interacting with upstream region of soybean ${\beta}-conglycinin$ gene- (대두 ${\beta}-conglycinin$ 유전자 발현의 전사 조절에 관한 연구 -(I) 대두 ${\beta}-conglycinin$ 유전자의 upstream 영역에 결합하는 대두 배 인자의 동정-)

  • Lee, Jeong-Yeon;Chung, Dong-Hyo;Kim, Woo-Yeon
    • Applied Biological Chemistry
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    • v.36 no.6
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    • pp.547-552
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    • 1993
  • Soybean nuclear extracts and S-100 were prepared to examine the soybean embryo factors which bind to the upstream region of soybean ${\beta}-conglycinin$ ${\alpha}'$ subunit gene. SEF3(soybean embryo factor 3), which is presumed to be a trans-acting factor for the expression of the gene, was detected in gel mobility shift assay using the DNA probe containing two AACCCA hexanucleotides. DNA probe containing CATGCAT or AACACA was used to find any other soybean embryo factor interacting with the upstream region of ${\beta}-Conglycinin$ ${\alpha}'$ subunit gene. It was found that there was no common DNA binding protein detected both in nuclear extracts and S-100. The relative levels of SEF3 binding activity both in nuclear extracts and S-100 of maturing soybean seeds were determined. SEF3 activity of nuclear extracts was first detected around 20 days after pollination and significantly increased around 32 days after pollination.

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Protease Properties of Protease-Producing Bacteria Isolated from the Digestive Tract of Octopus vulgaris (Octopus vulgaris의 장관으로부터 분리한 단백질 분해효소 생성 균주와 생성된 효소의 특성)

  • Liu, Qing;Ren, Pei;Piao, Meizi;Yang, Ji-Young
    • Journal of Life Science
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    • v.23 no.12
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    • pp.1486-1494
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    • 2013
  • A high protease-producing strain was isolated and identified from the digestive tract of octopus vulgaris by detecting a hydrolysis circle of protease and its activity. The strain was identified by morphology observation, biochemical experiments, and 16S rRNA sequence analysis. The protease obtained from the strain was purified by a three-step process involving ammonium sulfate precipitation, carboxy methyl-cellulose (CM-52) cation-exchange chromatography, and DEAE-Sephadex A50 anion-exchange chromatography. The properties of protease were characterized as well. The strain Bacillus sp. QDV-3, which produced the highest activity of protease, was isolated. On the basis of the phenotypic and biochemical characterization and 16S rRNA gene-sequencing studies, the isolate was identified as follows: domain: Bacteria; phylum: Firmicutes; class: Bacilli; order: Bacillales; family: Bacillaceae; and genus: Bacillus. The isolate was shown to have a 99.2% similarity with Bacillus flexus. A high active protease designated as QDV-E, with a molecular weight of 61.6 kDa, was obtained. The enzyme was found to be active in the pH range of 9.0-9.5 and its optimum temperature was $40^{\circ}C$. The protease activity retained more than 96% at the temperature of $50^{\circ}C$ for 60 min. Phenylmethylsulfonyl fluoride (PMSF) inhibited the enzyme activity, thus confirming that this protease isolated from Bacillus sp. QDV-3 is an alkaline serine protease. Metal ions, $Mn^{2+}$ and $Mg^{2+}$, were determined to enhance the protease activity, whereas $Ba^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ were found to inactivate the enzyme.

Identification of Agrobacterium tumefaciens from Soil and Transformation of Maize (토양으로부터의 Agrobacterium tumefaciens의 분리, 동정 및 옥수수의 형질전환에 이용)

  • 노광수;강봉중
    • KSBB Journal
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    • v.7 no.3
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    • pp.191-200
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    • 1992
  • Several strains of Agrobacterium tumefaciens were isolated from soil in the Taegu area and characterized to develop some useful vector systems for higher plant genetic engineering. The selected colonies had a unique form, and strains from the colonies were capable of tumor formation on the sunflower leaf surface. They had a large plasmid. The restriction analysis showed that they were another kinds of Ti plasmic compared with C58 and Ach5. The isolated strains were identified as the nopaline type and also as biovar 1 A. tumefaciens, according to their tumor morphology, blophyslcal and biochemical characteristics. One of the isolated strains, AK204 was transformed with binary vector (pGA642), having selectable marker (Kmr, Tcr). Furthermore, maize tissue cells were transformed by cocultivation with AK204/pGA642, and the transformants were selected on the selective medium and identified using PAGE patterns of their soluble proteins.

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Identification of Black Aspergillus Strains Isolated from Meju (메주에서 분리한 검은 Aspergillus 균주의 동정)

  • Hong, Seung-Beom;Kim, Dae-Ho;Kim, Seon-Hwa;Bang, Narae;Kwon, Soon-Wo
    • The Korean Journal of Mycology
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    • v.41 no.2
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    • pp.132-135
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    • 2013
  • Black Aspergillus is important fungus for oriental fermentation industry. Black Aspergillus was frequently isolated from Korean traditional Meju, a fermented soybean starting material for soy sauce and soybean paste. Thirty three strains were isolated from 98 finished Meju collected in various regions of Korea from 2008 to 2011, and 21 strains were isolated from in-process Meju at various farms from 2010 to 2011. The isolated black Aspergillus were identified using DNA sequences of partial ${\beta}$-tubulin and calmodulin genes. Of 54 black Aspergillus strains, 14 strains were identified as A. luchuensis and the others were composed of A. niger (n = 21), A. tubingensis (n = 10), and A. welwitschiae (n = 9).

Isolation and Characterization of Microorganisms for the Development of Fermentation Accelerator of Animal Manure (가축분뇨 발효제의 개발을 위한 미생물 분리 및 특성조사)

  • Kim, So-Young;Kim, Hong;Choi, Hee-Jung
    • KSBB Journal
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    • v.18 no.6
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    • pp.466-472
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    • 2003
  • Several microorganisms were isolated and characterized for the development of fermentation accelerator of animal manure. Firstly, 61 species were isolated from rice bran extract. Secondly, five strains of microorganisms were screened by the analysis of hydrolysis activities for organic compounds including protease, cellulase, amylase, and lipase. From a deodorization test for ammonia gas using the isolated strains, finally three bacterial strains were selected (NA 2, 12, 15). The selected strains, NA 2 and 15 were identified as Bacillus acidocaldarius and Planococcus sp. respectively. The media composition of key nutrients and pH for the mixed culture of the three selected strains were optimized using an experimental design method (response surface method) as follows : beef extract (4.59g/L), peptone (8.72g/L) and pH 6.3. Consequently, the isolated microorganisms seem to have potential applicability in the animal manure treatment.

The study on isolation of fibrinolytic bacteria from soybean paste (된장으로 부터 fibrin 용해 세균의 분리에 관한 연구)

  • Heo, Seok;Joo, Hyun-Kyu;Song, Ki-Bang;Lee, Si-Kyung
    • Applied Biological Chemistry
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    • v.42 no.1
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    • pp.6-11
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    • 1999
  • The bacteria which could hydrolyze the fibrin produced through the blood coagulation mechanism in the human body, were isolated from soybean paste. The KDO-13 strain was selected among the isolated bacteria as the best strain for fibrinolytic activity. It was spore forming and Gram positive. $C_{15:0}$ anteiso fatty acid, $C_{15:0}$ iso fatty acid and $C_{15:0}$ anteiso fatty acid were 47.7, 13.5 and 13.6%, respectively as major component among its cellular fatty acid composition. It showed the similarity of 57.7%, compared with standard strain. It was thus identified to be Bacillus atrophaeus according to Bergey's manual of systematic bacteriology and its fatty acid profiles of gas chromatography. The optimum culture temperature and pH were $37^{\circ}$ and 6 for the production of fibrinolytic enzyme by Bacillus atrophaeus KDO-13.

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Isolation and Characterization of the Strain Producing Angiotensin Converting Enzyme Inhibitor from Soy Sauce (간장으로부터 Angiotensin Converting Enzyme 활성 저해물질 생성 균주의 분리 동정)

  • 차명화;박정륭
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.4
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    • pp.594-599
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    • 2001
  • This study was attempted to isolate and identify the strain revealing high angiotensin converting enzyme (ACE) inhibitory activity from various soy fermented foods, i.e. meju, soybean paste and soy sauce. Forty-two strains with morphologically different characteristics were selected and the ACE inhibitory and proteolytic activities were examined. Of the strains tested, SS103 which was isolated from soy sauce showed the highest ACE inhibitory and proteolytic activities and was finally selected for further studies. The SS103 strain showed motility, rod form and ellipsoidal spores. The shape of colonies on the agar media was irregular, mucoidal and surface dull. The strain could grow under aerobic conditions of pH 5~9 and 10~$50^{\circ}C$. Main cellular fatty acid was $C_{15:0}$ anteiso, $C_{17:0}$ cis and $C_{17:0}$ iso, which was 33.9%, 18.8% and 16.5%, respectively. Based upon these morphological, biochemical and cultural properties, SS103 was identified as a Bacillus subtilis. Optimum cultural condition of Bacillus subtilis SS103 was pH 8.0, $37^{\circ}C$ and 48 hr.48 hr.

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Isolation of Bacteria Associated with Fresh Sponges in Lake Baikal (바이칼 호수에 서식하는 담수 스폰지 내 공생세균의 분리)

  • Cho, Ahn-Na;Kim, Ju-Young;Ahn, Tae-Seok
    • Korean Journal of Ecology and Environment
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    • v.47 no.spc
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    • pp.39-47
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    • 2014
  • Sponge in Lake Baikal is an unique organism. Microorganisms in sponges are assumed as precious resources for bioactive materials. For understanding the bacterial community in Baikalian sponges by cultivation, 92 strains of bacteria were isolated from lake water and 2 species of sponges, Baikalospongia sp. and Lubomirskia sp., Thirty five bacterial strains are isolated from ambient water near the sponge, 27 bacterial strains from Baikalospongia sp., 30 bacterial strains from Lubomirskia sp.. As a result, 78.3% and 57.6% of isolated bacterial strains has amylase and protease activity respectively, while strains with cellulose and lipase activities were 38.0% and 34.8%. By 16S rRNA sequence analysis of selected strains, 13 strains which were isolated from Baikalospongia sp. were belong to Pseudomonas spp.. Whereas, 14 strains which were isolated from Lubomirskia sp. were Pseudomonas spp., Buttiauxella agrestis, Pseudomonas fluorescens, Yersinia ruckeri, Bacillus spp., Paenibacillus spp., Bacillus thuringiensis, Bacillus simplex, Brevibacterium spp., Acinetobacter lwoffii. In culture media, Pseudomonas spp. dominance was supposed that according to allelophathy.

Identification of Calretinin-immunoreactive AII Amacrine Cells in the Brazilian Opossum (Monodelphis domestica) (브라질산 주머니쥐(Monodelphis domestica) 망막 내에서의 calretinin 면역반응성을 가지는 AII 무축삭세포의 동정)

  • Jeong, Se-Jin;Jeon, Chang-Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.19 no.2
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    • pp.271-277
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    • 2014
  • Purpose: The purpose of this study was to investigate the immunoreactivity of calretinin in Brazilian opossum (Monodelphis domestica) retina. Calcium-binding protein calretinin is known to play a key role in calcium-mediated signal transduction. Methods: Experiments have been performed by standard immunocytochemical techniques on retina of the Brazilian opossum. Results: Calretinin-immunoreactivity was exhibited within the horizontal subpopulations, AII amacrine and ganglion cell subpopulations in the Brazilian opossum retina. Especially, all calretinin-immunoreactive AII amacrine cells also expressed parvalbumin. Conclusions: Similar to other mammalian retinas, calretinin-immunoreactivity was also observed within the AII amacrine cells in the Brazilian opossum retina. Thus, calretinin can be a marker of AII amacrine cells in the Brazilian opossum retina.