• Title/Summary/Keyword: 31kDa

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경구투여 백신 후보물질로서의 Helicobacter pylori 외막 단백질의 조사

  • 박형배;최태부
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.129-136
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    • 1997
  • Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.

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Effect of Addition of Chitosan on Improvement for Shelf Life of Bread (키토산 첨가에 의한 식빵의 저장성 증진효과)

  • Lee, Hyun-Young;Kim, Seong-Mi;Kim, Jin-Young;Youn, Sun-Kyoung;Choi, Jung-Su;Park, Sun-Mee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.3
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    • pp.445-450
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    • 2002
  • This study was performed to evaluate the effect of chitosan addition on shelf life of bread. Four different molecular weights (MW) of chitosan, 1 kDa, 5 kDa, 30 kDa, and 120 kDa were used. The growth of spoilage bacteria was inhibited depending on MW and concentration of chitosan, and most inhibited conditions were 0.1% of 30 kDa and 120 kDa of chitosans. But the growth of yeast and mold showed very weak inhibition. This showed that yeast would grow normaly in the fermentation process of bread. Shelf-life of bread was improved depending on concentration of chitosans at WM 30 kDa and 120 kDa. Antioxidative effect of chitosan was increased with the larger molecular weight and the higher concentration during storage. The preservation of bread by adding chitosan was improved.

Molecular Cloning of a Gene Cluster for Phenanthrene Degradation from Pseudomonas sp. Strain DJ77 and Its Expression in Escherichia coli (Pseudomonas sp. strain DJ77로 부터 phenanthrene 분해 유전자군의 클로닝과 대장균에서의 발현)

  • 김영창;윤길상;신명수;김흥식;박미선;박희진
    • Korean Journal of Microbiology
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    • v.30 no.1
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    • pp.1-7
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    • 1992
  • We cloned a gene cluster encoding phenanthrene-degrading enzymes on a 6.8-kb Xhol fragment from the Pseudomonas sp. DJ77 chromosomal DNA into the vector pBLUESCRIPT SIC(+). The resultant clone, containing the recombinant plilsmid pHENX7, was able to convert 3-methylcatechol to a yellow mela-cleavage compound. Since the pHENX7R in which the DNA insert was cloned in the opposite orientation lacked extradiol dioxygenase activity. the direction of transcription was established. Four polypeptides, PhnC (24 kDa). PhnD (31 kDa), PhnE (34 kDa). and PhnF (15 kDa), were identified in E coli JM101 transformed with several pHENX7-derived plasmids. The locations and extents of ~ndividual genes were determined by subcloning. The gene order was phnC-phnD-phnE-phnF-phnG, and phnC, phnD, phnE, and phnG genes encoded glutathione S-transferase, mrta-cleavage compound hydrolase, extradiol dioxygenase, mera-cleavage compound dehydrogenase, respectively.

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Electrochemical Determination of Dopamine Based on Carbon Nanotube-Sol-Gel Titania-Nafion Composite Film Modified Electrode

  • Park, Ji-Ae;Kim, Byung-Kun;Choi, Han-Nim;Lee, Won-Yong
    • Bulletin of the Korean Chemical Society
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    • v.31 no.11
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    • pp.3123-3127
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    • 2010
  • A highly sensitive electrochemical detection method for dopamine (DA) has been developed by relying on a multiwalled carbon nanotube (CNT)-sol-gel titania-Nafion composite film modified glassy carbon (GC) electrode. The CNT-titania-Nafion/GC electrode exhibited excellent electrocatalytic activity towards DA. Therefore, the CNT-titania-Nafion/GC electrode showed improved voltammetric and amperometric responses for DA compared to those obtained with both titania-Nafion/GC and Nafion/GC electrodes. The CNT-titania-Nafion/GC electrode gave a linear response ($R^2$ = 0.999) for DA from $0.5\;{\mu}M$ to 0.5 mM with a detection limit (S/N = 3) of $0.1\;{\mu}M$ and a good sensitivity of 150 mA/M while other electrodes such as CNT-Nafion/GC, titania-Nafion/GC, and a bare GC gave a sensitivity of 89, 39, and 36 mA/M, respectively. Besides, the CNT-titania-Nafion/GC electrode displayed very fast response time within 2 s. The modified electrode showed good selectivity against ascorbic acid. The modified electrode showed good stability and reproducibility. The CNT-titania-Nafion/GC electrode was applied to the determination of DA in urine and serum samples.

Localization and isozyme patterns of phosphatase in Fibricola seoulensis (Fibricola seoulensis에서 phosphatase의 분포와 동위효소유형)

  • 김홍자;김창환
    • Parasites, Hosts and Diseases
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    • v.31 no.4
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    • pp.353-362
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    • 1993
  • The present study was carried out to investigate the localization and isozyme patterns of acid phosphatase and alkaline phosphatase in metacercariae and in adults of F. seoulensis by enzyme-histochemistry method and electrophoresis. Acidphosphatase showed a strong activity at pH 5 in the intestinal caecum of adults, but showed no reactions in the nonsubstrate control and in the inhibitor-treated control. Alkaline phosphatase showed a strong activity at pH 8 in the intestinal caecum and the tribocytic organ of adults, and in the intestinal caecum and in the genital anlagen of metacercariae. In non-denature PAGE, ten bands of protein fraction from the extracts of metacercariae and twenty-two bands from adults were detected. In denature PAGE, two protein bands having molecular weights of 192 kDa and 123 kDa were detected in the metacercariae, but absent from adult stage. In adults, protein fractions of 27.5 kDa, 24.5 kDa, 21.4 kDa, 18 kDa, 16 kDa and 15 kDa were detected. In non-denature PAGE, isozymes of acid phosphatase showed the most strong activity at pH 5, whereas no activity was shown at pH 2 and pH 7. One isozyme 85 kDa, 73 kDa and 62 kDa) in adults.

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Antibody Responses to Cryptosporidium Antigen in HIV-positive Patients in the Republic of Korea

  • Guk, Sang-Mee;Chai, Jong-Yil;Shin, Yung-Oh;Seo, Min
    • Parasites, Hosts and Diseases
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    • v.46 no.2
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    • pp.71-75
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    • 2008
  • The diagnosis of cryptosporidiosis has been carried out using coprologic techniques in the Republic of Korea. However, antibody responses to Cryptosporidium have rarely been studied. Serum antibodies from HIV-positive/oocyst-positive Korean patients recognized significantly 31 and 27 kDa antigens, and HIV-negative/oocyst-positive individuals clearly reacted to 15/17 kDa antigens. Compared with oocyst-positive cases, 18.7% and 75.8% of sera from HIV-positive patients reacted to 31 and 27 kDa antigens. Only 11.1% of HIV-negative individuals reacted to 15/17 kDa. Based on these findings, serum antibody responses were different between HIV-positive and HIV-negative individuals infected with Cryptosporidium, and it is suggested that HIV-positive patients are more frequently exposed to C. parvum compared to HIV-negative individuals.

Characterization of a Late Gene, ORF60 from Bombyx mori Nucleopolyhedrovirus

  • Du, Meng-Fang;Yin, Xin-Ming;Guo, Zhong-Jian;Zhu, Liang-Jun
    • BMB Reports
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    • v.39 no.6
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    • pp.737-742
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    • 2006
  • Open reading frame 60 of Bombyx mori nucleopolyhedrovirus (Bm60) is located between 56,673 and 57,479 bp in the BmNPV genome which encodes 268 amino acid residues with predicted molecular weight of 31.0 kDa. Bm60 and its homologues have been identified in 11 completely sequenced lepidopteran NPVs. The transcript of Bm60 was detected by RT-PCR at 18-72 h post-infection (p.i.), while the corresponding protein could be detected at 24-72 h p.i. in BmNPV-infected BmN cells by Western blot analysis using a polyclonal antibody against Bm60. The expression of Bm60 was inhibited in the presence of Ara-c, an inhibitor of viral DNA synthesis. These results together indicated that Bm60 was a late gene. The size of Bm60 product was found to be a 31 kDa in BmNPV-infected BmN cells, consistent with predicted molecular weight. Immuno-fluoresence analysis showed that the Bm60 product was first detected in the cytoplasm at 24 h p.i and also located in nucleus during later infection. In conclusion, the available data suggest that Bm60 is a functional ORF of BmNPV and encodes a 31kDa protein expressed in the later stage of infection cycle.

Alterations in the Level of Fibronectin and its Receptors during Chick Myoblast Differentiation (계배 근원세포 분화에 따른 Fibronection의 수준과 그 수용체의 변화)

  • 정창용;강만식
    • The Korean Journal of Zoology
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    • v.31 no.2
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    • pp.95-103
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    • 1988
  • Alterations in the amount of fibronectin during chick myogenesis were investigated. The amount of fibronectin as measured by immunoblotting was found to decrease during the process. As a first step in answering the precise mode of change in the level of ibronedin during myogensis, the interaction of 28,000 dalton(28 kDa) amino terminal fragment of fibronectin as well as 85,000 dalton (85 kDa) fragrxient with myoblasts was examined. The specific binding of 125 l-28 kDa fragment to myoblasts was time-dependent and reached a maximum within 60 min. Unlabelled 28 kDa fragment inhibited the binding of 125 I-28 kDa fragment, whereas 85 kDa fragment containing adhesion promoting activity did not inhibit it. This finding suggests that the 28 kDa fragment interacts with the matrix assembly receptors but not with the cell adhesion receptors. Accordingly, the decrease in the level of fibronectin is likely to correlate with the fall of fibronectin receptors on the myoblasts.

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Phosphorylation of a 66 kDa Protein, a Putative Protein Kinase C Substrate, is Related to Chondrogenesis of Chick Embryo Mesenchymes In Vitro

  • Lee, Sun-Ryung;Sonn, Jong-Kyung;Yoo, Byung-Je;Lim, Young-Bin;Kang, Shin-Sung
    • BMB Reports
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    • v.31 no.4
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    • pp.350-354
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    • 1998
  • To understand the role of protein kinase C (PKC) in the regulation of chondrogenesis, we examined proteins which are phosphorylated by PKC. Stage 23/24 chick embryo wing mesenchymes were micromass-cultured to induce chondrogenesis and cell extracts were phosphorylated in a condition that activates PKC. Several proteins including 63 and 66 kDa proteins were phosphorylated. The 66 kDa protein was phosphorylated only in the presence of phorbol 12-myristate 13-acetate (PMA) and phosphatidylserine CPS), and the phosphorylation was almost completely diminished by bisindolylmaleimide, a PKC inhibitor. In addition, partially purified PKC increased the phosphorylation of the 66 kDa protein. Treatment of cultures with lysophosphatidylcholine (LPC) promoted chondrogenesis and phosphorylation of 66 kDa protein, while PMA and thymeleatoxin inhibited both of the two events. Our results suggest that the 66 kDa protein is a putative substrate of PKC, and phosphorylation of the 66 kDa protein, probably by $PKC\alpha$ is required for chondrogenesis.

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Identification of Allergens of Dermatophagoides farinae on Canine Atopic Dermatitis in Korea (국내 개에서 발생한 아토피성 피부염에서 Dermatophagoides farinae의 항원 확인)

  • Cho, Na-Young;Park, Seong-Jun
    • Journal of Veterinary Clinics
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    • v.31 no.6
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    • pp.490-494
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    • 2014
  • Dermatophagoides farinae plays important role in the pathogenesis of canine atopic dermatitis as environmental allergens. Also, many studies revealed that D. farinae was the main causative allergen for Korean dogs with atopic dermatitis. To identify major allergens of D. farinae in Korean atopic dogs allergic to D. farinae by immunoblot using commercial allergenic extracts, 26 dogs from two groups were enrolled in the study. Control group consists of 10 dogs with no clinical signs of disease and atopic group consists of 16 dogs diagnosed as atopic dermatitis. Sera from Korean dogs with atopic dermatitis showed six allergens of D. farinae extract by procedure of immunoblot. The molecular weights of identifying protein bands were 177, 109, 75, 44, 27, 15 kDa. The major allergens showing reactivity with greater than 50% of atopic dogs were detected at approximately 44, 109 and 177 kDa. Subsequent investigations will be carried out to verify the identity of the allergens detected in this study.