• Title/Summary/Keyword: GP5

Search Result 544, Processing Time 0.026 seconds

Studies on the cloning gp50 and gp63 genes of Pseudorabies virus(Shope strain) (Pseudorabies virus의 gp50과 gp63 유전자 클로닝에 관한 연구)

  • Kweon, Chang-hee;Song, Jae-young;Kim, Byoung-han;Lee, Jung-bok;Lee, jae-chin;An, Soo-hwan;Lee, Yong-soon;Susumu, Maeda
    • Korean Journal of Veterinary Research
    • /
    • v.31 no.3
    • /
    • pp.311-318
    • /
    • 1991
  • The DNA fragment representing for Pseudorabies gp50 and gp60(Shope) was cloned by recombinant techniques. The viral DNA was extracted from the infected cells and digested with Bam HI. The 6.8 Kb of Bam HI fragment was isolated from agarose gel and further digested with Nde I followed by Klenow treatment. The blunt ended 4.9Kb fragment was cloned into pTZ18R plasmid vector. The upstream region of gp50 was further manipulated to remove its 5' promoter region and create EcoRl site for possible eukaryotic expression system. The result of partial sequencing of cloned DNA indicated that Shope strain showed 95% homology with gp50 of Rice strain.

  • PDF

Preparation and Characterization of Anti-GP73 Monoclonal Antibodies and Development of Double-antibody Sandwich ELISA

  • Li, Qi-Wen;Chen, Hong-Bing;Li, Zhi-Yang;Shen, Peng;Qu, Li-Li;Gong, Lai-Ling;Xu, Hong-Pan;Pang, Lu;Si, Jin
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.16 no.5
    • /
    • pp.2043-2049
    • /
    • 2015
  • Background: Serum Golgi protein 73 (GP73) as a novel and potential marker for diagnosing hepatocellular carcinoma (HCC) have been found to be elevated in HCC patients and associated with clinical variables representing tumor growth and invasiveness. The aim of this study was to prepare a pair of monoclonal antibodys (mAbs) against GP73 and develop a newly designed double-antibody sandwich enzyme-linked immunosorbent assay (s-ELISA), which would be used in the detection of serum GP73 (sGP73) as well as in the diagnosis of HCC. Materials and Methods: Produced by prokaryotic expression, the purified recombinant GP73 (rGP73), produced by prokaryotic expression, was used to immunize the Balb/c mice. Two hybridoma cell lines against GP73 were obtained by fusing mouse Sp2/0 myeloma cells with spleen cells from the immunized mice. The titers of anti-GP73 mAb reached 1:243,000. Western blotting analysis and Immunohistochemistry staining revealed that anti-GP73 mAb could recognize GP73 protein. The double-antibody s-ELISA was successfully established and validated by 119 HCC and 103 normal serum samples. Results: showed that the detection limit of this method could reach 1.56 ng/ml, and sGP73 levels in HCC group (mean=190.6 ng/ml) were much higher than those of in healthy controls (mean=70.92 ng/ml). Conclusions: Results of our study not only showed that sGP73 levels of HCC patients were significantly higher than those of healthy controls, but also indicated that the laboratory homemade anti-GP73 mAbs could be the optimal tool used in evaluating sGP73 levels, which would provide a solid foundation for subsequent clinical applications.

Screening for Chemosensitizers from Natural Plant Extracts through the Inhibition Mechanism of P-glycoprotein

  • Ahn, Hee-Jeong;Song, Im-Sook
    • Journal of Pharmaceutical Investigation
    • /
    • v.40 no.5
    • /
    • pp.269-275
    • /
    • 2010
  • P-gp plays a critical role in drug disposition and represents a mechanism for the development of multidrug resistance. Flavonoids, a major class of natural compounds widely present in foods and herbal products, have been shown to inhibit P-gp. Therefore, the aim of this study was to identify new candidate chemosensitizers by screening various plant extracts. The ability of natural plant extracts to inhibit P-gp activity was assessed by measuring cellular accumulation of calcein AM, daunorubicin and vincristine in P-gp overexpressing MDCKII-MDR1 cells. Among more than 800 plant extracts, eight were found to inhibit P-gp activity. Curcuma aromatica extract produced greatest inhibition, followed by Curcuma longa and Dalbergia odorifera extracts. Extracts of Aloe ferox, Curcuma zedoariae rhizome, Zanthoxylum planispinum, and Ageratum conyzoides showed moderate inhibitory effects. Curcumin and quercetin exhibited similar inhibition of P-gpmediated efflux of daunorubicin and vincristine, and flavones had a lesser effect. When chemosensitizing effect was evaluated by measuring daunorubicin sensitivity to MDCKII-MDR1 cells in the presence of natural plant extracts, Curcuma aromatica showed the most potent chemosensitizing effect based on daunorubicin cytotoxicity. In conclusion, natural plant extracts such as Curcuma aromatica can potently inhibit P-gp activity and may have potential as a novel chemosensitizers.

Effects of PEG Priming Treatment on Germination and Seedling Growth of Onion Seed(Allium cepa L.)

  • Lee, Sheong-Chun;Ahn, Chan-Young
    • Plant Resources
    • /
    • v.2 no.2
    • /
    • pp.88-95
    • /
    • 1999
  • These experiments were conducted to evaluate the variability of seed germination and seedling growth with different levels of polyethylene glycol(PEG 6,000) solution in onion seed. Average germination percentage of seed primed in PEG solution with 1.00 and 0.75 MPa was higher than control, and that of seed primed in 1.50MPa was lower than unprimed control. Germination percentage(GP) of seed primed for 5 days was highest, and as the primed days become long, the GP was decreased. The GP of airation seed during the primed was higher than that of unairation seed, about 5% , respectively. The GP of washed seed after primed was higher than that of unwashed seed, but that of redried seed after primed was lower than that of the others. The highest GP cultivar was Chunjoogoohyung and the lowest GP cultivar was Seouldego in unredried seed after primed, but Chunjoojoonggo was highest and Jungpoonwhang was the lowest cultivar in redried seed after primed. As the PEG concentration increased, the seedling length(SL) was shortened, and seed primed for 15 days was longer than other treatments. The SL of primed seed was similar to GP. The SL of washed seed after primed was longer than that of others, but that of redried seed after primed was shortest among the others. The SL of Chunjoojoonggo and Nongwoodego was longest and Seouldego was shortest among the cultivars in unredried seed after primed, but that of Chunjoogoohyung and Chunjoojoonggo was longest and Seouldego was the shortest cultivars in redried seed after primed.

  • PDF

Effect of Gaeddongssuk (Artemisia annua L.) Powder on Quality and Shelf Stability of Emulsion Sausages during Refrigerated Storage

  • Ham, Hyoung-Joo;Kang, Geun-Ho;Choi, Yun-Sang;Jeong, Tae-Jun;Hwang, Ko-Eun;Kim, Cheon-Jei
    • Food Science of Animal Resources
    • /
    • v.36 no.5
    • /
    • pp.601-611
    • /
    • 2016
  • The objective of this study was to evaluate effects of Gaeddongssuk powder (GP) on quality characteristics and shelf stability of emulsion sausages during storage. Proximate composition properties showed no significant differences in all treatment (p>0.05). Control showed the highest cooking loss while the treatment with GP showed decreased cooking loss depending on increasing GP content (p<0.05). Apparent viscosity of batter was increased as the amount of GP increased, whereas hardness of emulsion sausages was decreased with increasing GP level. In sensory evaluation, emulsion sausage with 0.1% GP resulted in the highest score in overall acceptability. The pH values of all treatments decreased at the early storage stage, followed by gradual increase. The lightness and redness of treatments were decreased when the level of GP was increased. However, the yellowness of sausages with GP were higher than that of control (p<0.05). The addition of GP inhibited lipid oxidation of emulsion sausages during storage depending on its level. The aerobic bacteria population and VBN was unaffected by addition of GP during the storage (p>0.05). Therefore, Gaeddongssuk powder up to 0.1% has a potential as a natural antioxidant for meat products because it can inhibit lipid oxidation of sausages without decreasing their sensory properties.

Development of Microbial Augmentation for the Treatment of Recalcitrant Industrial Wastewater Containing Chlorinated Organic Compounds (유기염소계 난분해성 산업폐수의 처리를 위한 미생물제제의 개발)

  • Lee, Hyun Don;Im, Seong Won;Suh, Hyun-Hyo
    • Journal of Life Science
    • /
    • v.24 no.8
    • /
    • pp.887-894
    • /
    • 2014
  • The screening of the microorganisms degrading chlorinated organic compounds such as PCP (pentachlorophenol) and TCE (trichloroethylene) was conducted with soil and industrial wastewater contaminated with various chlorinated organic compounds. Isolates (GP5, GP19) capable of degrading PCP and isolates (GA6, GA15) capable of degrading TCE were identified as Acetobactor sp., Pseudomonas sp., Arthrobacer sp., Xanthomonas sp. and named Acetobacter sp. GP5, Pseudomonas sp. GP19, Arthrobacer sp. GA6 and Xanthomoas sp. GA15, respectively. The microbial augmentation, OC17 formulated with the mixture of bacteria including isolates (4 strains) degrading chlorinated organic compounds and isolates (Acinetobacter sp. KN11, Neisseria sp. GN13) degrading aromatic hydrocarbons. Characteristics of microbial augmentation OC-17 showed cell mass of $2.8{\times}10^9CFU/g$, bulk density of $0.299g/cm^3$ and water content of 26.8%. In the experiment with an artificial wastewater containing PCP (500 mg/l), degradation efficiency of the microbial augmentation OC17 was 87% during incubation of 65 hours. The degradation efficiency of TCE (300 uM) by microbial augmentation OC17 was 90% during incubation of 50 hours. In a continuous culture experiment, analysis of the biodegradation of organic compounds by microbial augmentation OC17 in industry wastewater containing chlorinated hydrocarbons showed that the removal rate of COD was 91% during incubation of 10 days. These results indicate that it is possible to apply the microbial augmentation OC17 to industrial wastewaters containing chlorinated organic compounds.

Induction of Apoptosis and Growth-Inhibition by Thymoquinone in ACHN and GP-293 Cell Lines in Comparable with Cis-Platinum

  • Shahraki, Samira;Mohebbati, Reza;Shafei, Mohammad Naser;Mahmoudi, Mahmoud;Hosseinian, Sara;Parhizgar, Soghra;Yazd, Zohreh Naji Ebrahimi;Heravi, Nazanin Entezari;Abadi, Reza Nejad Shahrokh;Khajavirad, Abolfazl
    • Journal of Pharmacopuncture
    • /
    • v.22 no.3
    • /
    • pp.176-183
    • /
    • 2019
  • Objective: In the current work, we investigated the cytotoxic and apoptotic effects of Thymoquinone (TQ), an active compound of Nigella sativa (N. sativa) and Cis-platinum, on normal renal epithelial (GP-293) and human renal adenocarcinoma cell lines (ACHN). Methods: GP-293 and ACHN cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% Fetal bovine serum (FBS) and 1% penicillin plus streptomycin antibiotic. The MTT assay was used for cellular viability assessment. Viability of cells was observed using inverted light microscope 24, 48 and 72 h after exposure of the cells to various concentrations of TQ (1, 2.5, 5, 10, 50 and $100{\mu}g/ml$) and Cis-platinum (0.5, 1, 1.5, 2, 3, 6 and $12.5{\mu}g/ml$). Moreover, apoptosis was analyzed with a flow-cytometry method. The untreated cells were considered as control group. Results: Morphological changes such as reduced cell number and increased intercellular distance and reduced cell viability in ACHN and GP-293cell lines were observed in both TQ and Cis- platinum groups; however, Cis-platinum had greater effect on ACHN cell line than GP-293 cell line. In addition, GP-293 cell line was more sensitive to TQ compared to ACHN cell line. Furthermore, TQ and Cis-platinum had apoptotic effects on both ACHN and GP-293 cell lines. Conclusion: Our findings demonstrated that TQ and Cis-platinum had cytotoxic and apoptotic effects on both cell lines, However, GP-293 cell line was more sensitive to TQ. Additionally, Cis-platinum was more effective on ACHN cell line than on GP-293 cell line.

Antimutagenic and Anticancer Effects of Glycoprotein and Chondroitin Sulfates from Sea Cucumber(Stichopus japonicus) (해산 극피동물 중의 당단백질의 특성과 이용 II. 해삼당단백질과 황산콘드로이친의 항돌연변이 및 항암효과)

  • 류홍수;문정혜;양훈석;서재수
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.27 no.2
    • /
    • pp.350-358
    • /
    • 1998
  • The antimutagenic and anticancer activities of glycoprotein(GP) and chondroitin sulfate(CS) from sea cucumbers were studied using Ames mutagenicity test and human cancer cells culture test. The GP's inhibitory effect toward aflatoxin B1(AFB1) and 3, 2'-dimethyl-4-amino-biphenyl(DMAB) increased with the higher added concentrations up to 5% level(w/w) regardless fractionation methods. The GP from sea cucumbers through DEAE-cellulose column chromatography showed an inhibitory effect ranged from 84 to 98%, and the maximum antimutagenicities resulted in red sea cucumber with 98% (AFB1) and 95% (DMAB). But 5% level of CS from various sea cucumbers had an inhibitory effect toward those both indirect mutagens ranged from 79% to 85%. However, in case of direct mutagens(N-methyl-N'-nitro-N-nitrosoguanidine, MNNG and 4-nitroquinoline-1-oxide, 4-NQO), the GP's inhibitory effect was 55∼78% and the CS had a low inhibitory effect(58∼70%) at the added level of 5%. The GP from sea cucumbers exhibited the strong inhibitory effects with 89∼95% and 82∼92% on the growth of HT-29 human crcinoma cells and AZ-521 human gastric cancer cells (at 5% level).

  • PDF

Micro-computed tomographic evaluation of single-cone obturation with three sealers

  • Sahar Zare;Ivy Shen;Qiang Zhu;Chul Ahn;Carolyn Primus;Takashi Komabayashi
    • Restorative Dentistry and Endodontics
    • /
    • v.46 no.2
    • /
    • pp.25.1-25.12
    • /
    • 2021
  • Objectives: This study used micro-computed tomography (µCT) to compare voids and interfaces in single-cone obturation among AH Plus, EndoSequence BC, and prototype surface pre-reacted glass ionomer (S-PRG) sealers and to determine the percentage of sealer contact at the dentin and gutta-percha (GP) interfaces. Materials and Methods: Fifteen single-rooted human teeth were shaped using ProTaper NEXT size X5 rotary files using 2.5% NaOCl irrigation. Roots were obturated with a single-cone ProTaper NEXT GP point X5 with AH Plus, EndoSequence BC, or prototype S-PRG sealer (n = 5/group). Results: The volumes of GP, sealer, and voids were measured in the region of 0-2, 2-4, 4-6, and 6-8 mm from the apex, using image analysis of sagittal µCT scans. GP volume percentages were: AH Plus (75.5%), EndoSequence BC (87.3%), and prototype S-PRG (94.4%). Sealer volume percentages were less: AH Plus (14.3%), EndoSequence BC (6.8%), and prototype S-PRG (4.6%). Void percentages were AH Plus (10.1%), EndoSequence BC (5.9%), and prototype S-PRG (1.0%). Dentin-sealer contact ratios of AH Plus, EndoSequence BC, and prototype S-PRG groups were 82.4% ± 6.8%, 71.6% ± 25.3%, and 70.2% ± 9.4%, respectively. GP-sealer contact ratios of AH Plus, EndoSequence BC, and prototype S-PRG groups were 65.6% ± 29.1%, 80.7% ± 25.8%, and 87.0% ± 8.6%, respectively. Conclusions: Prototype S-PRG sealer created a low-void obturation, similar to EndoSequence BC sealer with similar dentin-sealer contact (> 70%) and GP-sealer contact (> 80%). Prototype S-PRG sealer presented comparable filling quality to EndoSequence BC sealer.

Molecular and Biochemical Studies on the DNA Replication of Bacteriophage T7: Functional Analysis of Amino-terminal Region of Gene 2.5 Protein

  • Kim, Young-Tae;Lee, Sung-Gu;Kim, Hak-Jun
    • BMB Reports
    • /
    • v.28 no.6
    • /
    • pp.484-489
    • /
    • 1995
  • The product of bacteriophage T7 gene 2.5 is a single-stranded DNA binding protein and plays an important role in T7 DNA replication, recombination, and repair. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth (Kim and Richardson, 1993). The C-terminal truncated gene 2.5 protein ($GP2.5-{\Delta}21C$) cannot substitute for wild-type gene 2.5 protein in vivo; suggesting that the C-terminal domain of gene 2.5 protein is essential for protein-protein interactions (Kim and Richardson, 1994; J. Biol. Chem. 269, 5070-5078). Truncated gene 2.5 proteins lacking 19 residues ($GP2.5-{\Delta}19N$) and 39 residues ($GP2.5-{\Delta}39N$) from the amino-terminal domain were constructed by in vitro mutagenesis. $GP2.5-{\Delta}19N$ can support the growth of T7 phage lacking gene 2.5 while $GP2.5-{\Delta}39N$ cannot substitute for wild-type gene 2.5 protein in vivo; however, its ability to bind to single-stranded DNA is not affected. These results clearly demonstrate that the 20~39 amino-terminal region of gene 2.5 protein is required for T7 growth in vivo but may not be involved in DNA binding activity.

  • PDF