• Title/Summary/Keyword: blot

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Effect of Water Extracts from Thesium chinense Tunczaninov and Prunella vulgaris L. on Aromatase and Cyclooxygenase Activities (하고초 열수추출물이 Aromatase와 Cyclooxygenase 활성에 미치는 영향)

  • Nam, Kyung-Soo;Shon, Yun-Hee
    • Korean Journal of Pharmacognosy
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    • v.35 no.2 s.137
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    • pp.147-151
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    • 2004
  • Water extracts from Thesium chinense Tunczaninov (TCTW) and Prunella vulgaris L. (PVW) were tested for aromatase and cyclooxygenase activities. TCTW and PVW were capable of suppressing aromatase in a human placenta microsomal assay. PVW was shown to be more effective than TCTW in the suppression of aromatase activity. TCTW significantly inhibited cyclooxygenase-2 (COX-2) activity at the concentration of 0.25 (p<0.05), 0.5 (p<0.01) and 2.5 mg/ml (p<0.005). PVW also inhibited COX-2 activity in a dose-dependent manner in a concentration range of $0.05{\sim}2.5\;mg/ml$. The expression of COX-2 was inhibitied by TCTW and PVW in western blot analysis. These results suggest that TCTW and PVW may have breast cancer chemopreventive potentials by inhibiting aromatase and cyclooxygenase activities.

Cloning and characterization of the psbA Gene from Panax ginseng(Characterization of the psbA Gene from P. ginseng)

  • Lee, Won-Kyu;Tae, Gun-Sik
    • Journal of Photoscience
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    • v.10 no.3
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    • pp.245-249
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    • 2003
  • The psbA gene of photo system II was cloned and characterized from the P. ginseng chloroplast. The psbA gene is composed of 1,062 nucleotides. The overall amino acid sequence shows 99% and 98% identities to dicots and monocots of higher plants, respectively. Southern blot analysis revealed that a single copy of the psbA gene existed in the chloroplast genome. Northern blot analysis of the in vivo accumulation of the psbA transcript, after being grown under the different intensities (5%, 10%, 20%, and 100%) of daylight, indicated that the steady-state level of the psbA transcript was not significantly affected by light intensity.

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The Effect of $\delta$-endotoxin of Bacillus thuringiensis on the Protein Synthesis of the Several Tissues of Hyphantria cunea Drury (Bacillus thuringiensis 내독소단백질이 미국흰불나방 조직의 단백질 합성에 미치는 영향)

  • 전향미;강석권;서숙재
    • Journal of Sericultural and Entomological Science
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    • v.37 no.1
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    • pp.68-73
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    • 1995
  • The toxic effect of Bacillus thuringiensis var, kurstaki $\delta$-endotoxin was determined in the midgut, fat body and Malpighian lobules from larvae of Hyphantria cunea using the au-toradiography and Western blot methods. Bt $\delta$-endotoxin crystals inhibited protein synthesis and elongation factor-2 activity of all tissues tested.

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Different level of tumor necrosis factor-α expression after administration of silk sericin fraction in RAW264.7 cells

  • Kim, Dae-Won;Jo, You-Young;Kweon, HaeYong;Kim, Seong-Gon
    • International Journal of Industrial Entomology and Biomaterials
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    • v.41 no.1
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    • pp.1-5
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    • 2020
  • Tumor necrosis factor-α (TNFα) is a representative marker for inflammation. Silk sericin is known as mild TNFα inducer. The purpose of this study was to compare the level of TNFα among different fractions of silk sericin. Silk sericin was extracted from cocoon and separated it by molecular weight. Each fraction was applied to RAW264.7 cells. The level of TNFα was evaluated by western blot and ELISA assay. In results, the level of TNFα was increased as time-dependent manner. Higher molecular weight fraction of sericin induced higher amount of TNFα than lower molecular weight fraction. In conclusion, different molecular weight fraction of sericin induced TNFα differently.

Characterization of ORF39 from Helicoverpa armigera Single-nucleocapsid Nucleopolyhedrovirus, the Gene Containing RNA Recognition Motif

  • Xu, Hai-Jun;Liu, Yan-He;Yang, Zhang-Nv;Zhang, Chuan-Xi
    • BMB Reports
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    • v.39 no.3
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    • pp.263-269
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    • 2006
  • In the genome of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus, open reading frame 39 (Ha39) is the only gene predicted to encode an RNA recognition protein. Computer analysis revealed that Ha39 homologues were found in 15 NPVs, but not in GVs. Its transcripts were detected from 3 through 72 hours post infection (h p.i.) using RT-PCR and Northern blot analysis. The protein was detected in infected-cell lysates from 6 h p.i. Western blot assay of ODV and BV preparations revealed that Ha39 encodes a structural protein associated with BVs. Additionally, immunofluorescence microscopy demonstrated that the protein was present within cytoplasm in virus-infected cells, but not in the nuclear region.

Purification and Partial Immuno - Characterization of Boar Sperm Proteinase Sperminogen

  • YiLee, S.H.
    • BMB Reports
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    • v.30 no.6
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    • pp.448-452
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    • 1997
  • Polyclonal antibody of the boar sperminogen was used to characterize the boar sperm proteinase sperminogen. Boar sperminogen was purified from the acid extracts of the washed epididymal spermatozoa by gel filtration through a Sephadex G-100 column. followed by preparative SDS-PAGE. The sperminogen band was sliced out and was eluted from the gel matrix. The purified sperminogen was used to produce the polyclonal antibody of the boar sperminogen. When characterized on a Western blot, the final preparation of sperminogen appeared as a homogenous protein with a molecular weight of 32 kDa. The relative migration of sperminogen was distinctly different from the major components of the proacrosin-acrosin system as well as all the observable proacrosin activation by-products detected on the Western blot. The sperminogen antibody, however. cross-reacted with the proacrosin-acrosin system.

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Inositol(1,4,5)triphosphate 3-Kinase의 유전자 재조합과 CCL39 Hamster Lung Fibroblasts에서 발현

  • 김재웅;최관용
    • The Korean Journal of Food And Nutrition
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    • v.9 no.2
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    • pp.123-136
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    • 1996
  • IPSKCDNA gene(1.8 kbp) encoding rat brain IP3K enzyme contained Not I restric site in open reading frame. The Not I sequence, GCGGCCGC, was converted to GCAGCCGC by site-directed mutagenesis. The mutated IP3KcDNA was digested with EcoR I and ligated with EcoR I-restricted psp72·Not2 vector. The resulting psp72 · Not2-IP3KCDNA was digested with the Not I restriction enzyme and then subcloned into the Not I -digested PZIP · NeoSV(X) mammalian expression vector. The PZIP · NeoSV(X) -IPSKCDNA was transfected into CCL39 hamster lung fibroblast cells. The efficiency of the expressed IPSKCDNA gene was significantly higher than expected generally, not only a mean 5-fold increase in the amount of enzyme, but also 16-fold increase in enzyme activity from tractsfected CCL39 cells by the method of Western blot using anti-lP3K antibodies. Both distribution of IPSK in various rat tissues and biochemical properties were discussed.

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Immunological Characterization of Bacillus thuringiensis Antigens (Bacillus thuringiensis 항원들의 면역학적 분석)

  • Jung, Jae-Deuk;Park, Jung-Sun;Jo, Young-Soo;Hong, Soon-Bok;Lee, Hyung-Hoan;Cho, Myung-Hwan
    • Microbiology and Biotechnology Letters
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    • v.23 no.1
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    • pp.110-117
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    • 1995
  • This study was carried out to immunologically characterize Bacillus thuringiensis (B.t) antigens. Protein patterns of ultrasonicated- antigens of B. thuringiensis subspecies using SDS- PAGE revealed marked similarities among all the strains analyzed except for the difference between quantative variations of bands and some protein antigens. The comparison of the protein patterns showed that the protein antigen of 45 kilodalton (kd) was common in 11 strains and that the difference between B. thuringiensis subsp. canadensis and galleriae was noticed in quantative variations of bands despite of ambiguous serogrouping, suggesting a useful method for identification. All strains examined showed similar antigenic patterns in SDS-PAGE, while immunodominant bands differed in antigenic reactivity in western blot using polyclonal antibodies. Polyclonal antibody to B. thuringiensis subsp. thuringiensis and israelensis in indirect immunofluorescence assay reacted with flagella and cell surface antigens. The present study indicates that SDS-PAGE and western blot analysis may be used as tools for differentiation and identification of B. thuringiensis subspecies.

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Identification of Prevotella intermedia ATCC 25611 Using Pi29-L DNA Probe. (Pi29-L DNA 프로브를 이용한 Prevotella intermedia ATCC 25611의 동정)

  • 국중기;백동헌
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.205-209
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    • 2003
  • Recently, we introduced a new method for rapid screening of bacterial species- or subspecies-specific DNA probes, named “inverted dot blot hybridization screening method”. We then applied this method to develop species- or strain- specific DNA probes for Prevotella intermedia and Prevotella nigrescens. In those studies, among 96 candidate DNA probes which were screened by the new method, 5 probes were confirmed as being putatively strain-specific : 3 probes for P. nigrescens 9336 (ATCC 33563), one for each p. intermedia ATCC 25611 and one for P. nigrescens G8-9K-3 (ATCC 49046). In the present study, we evaluated by Southern blot analysis a DNA probe Pi29-L, one of the 96 candidate probes described above, whether it is specific for the strain ATCC 25611 off. intermedia. Our data show that the probe Pi29-L is potentially P. intermedia ATCC 25611-specific, which can be useful for the detection and identification of the strain, particularly in maintenance of the strain.

Screening for Inhibitor of c-myc Expression and Identification of Isolate No.2303

  • Chung, Ji-Hyung;Yeo, Ick-Hyun;Oh, Doo-Whan;Moon, Soon-Ok
    • Journal of Microbiology and Biotechnology
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    • v.5 no.5
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    • pp.264-268
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    • 1995
  • Sulforhodamine B(SRB) assay was performed on the human lung carcinoma, A549 cell line to screen soil microorganisms for production of anti-cancer agent. Among 4, 265 microorganisms, 45 isolates were selected for their cytotoxicity and tested for their effects on the expression of c-myc by RNA slot blot and Northern blot analysis resulting in selection of No.2303 isolate. This No.2303 was identified as Streptomyces sp. by ISP classification and the chemotaxonomic analysis method. NO.2303 inhibited the expression of cmyc in Col0320 DM and A549 cell lines. The culture extract of No. 2303 also inhibited the progression of the cell cycle of Go in NIH 313 cells, implying that the extract also inhibited the expression of c-myc in NIH 313 cell.

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