• Title/Summary/Keyword: S glycoprotein

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Expression of Pseudorabies Virus Glycoproteins gB, gC and gD using Insect Expression System

  • Yun, Bit Na Rae;Gwak, Won Seok;Lee, Ji Hoon;Woo, Soo Dong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.35 no.2
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    • pp.118-122
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    • 2017
  • Porcine pseudorabies virus (PRV) causes the Aujeszky's disease (AD) which is economically important disease in the swine industry worldwide. Killed or live vaccines have been used to control this disease, but their efficacy and side effects remain problems to be solved. To solve these problems, in this study, production of recombinant PRV glycoprotein gB, gC and gD was investigated in insect expression system. Glycoprotein gB, gC and gD are regarded as the major immunogenic antigens in PRV. Abundant production and immunogenicity of glycoprotein gB, gC and gD were confirmed by SDS-PAGE and Western blot analysis, respectively. Optimal infection dose and time were also determined for the production of each recombinant PRV glycoprotein. Confirmation of glycosylation of recombinant gB, gC and gD suggested their usefulness as antigens for the development of diagnosis kit or vaccines for Aujeszky's disease.

Studies on the Compositon of Protein and lycoprotein in Sarcopiasmic Reticulum of Skeletal Muscle (근소포체의 단백질 및 당단백질 조성에 관한 연구)

  • 박영철
    • The Korean Journal of Zoology
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    • v.33 no.2
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    • pp.191-199
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    • 1990
  • Sarcoplasmic reticulum subfractions were isolated from rabbit sarcoplasmic reticulum vesicles using ultracentrifugation in a continuous sucrose gradient (12.5% 50%) after French pressure treatment. And proteins in sarcoplasmic reticulum were detected by SDS-polyacrylamide gel electrophoresis and glycoproteins were identified through the reaction with 1251-concanavalin A.The electrophoresis showed that sarcoplasmic reticulum contained predominantly $Ca^2$+-AThase and calsequestrin along with high affinity calcium binding protein, intrinsic glycoprotein 160 Kd, 94 Kd, 80 Kd, 38 Kd, 34 Kd and 24 Kd proteins. Among these, the protein of about 80 Kd which has been known as one of heat shock proteins was especially enriched in the terminal cistemae of sarcoplasmic reticulum. Meanwhile, autoradiogram of 125 I-concanavalin A bound to the stained gels showed the distribution of glycoproteins which included 160 Kd glycoprotein, 94 Kd glycoprotein, calsequestrin and intrinsic glycoprotein Among these, the protein of about 160 Kd was especially enriched in longitudial sarcoplasmic reticulum and T-tubule, and the protein of about 94 Kd which has been known as one of glucose-regulated proteins was also enriched in T-tubule and sharply reduced in terminal cistemae.

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Physicochemical Characteristics of Carrot Glycoprotein (당근 당단백질의 물리화학적 특성)

  • Lee, Mi-Jin;Jang, Boo-Sik;Jeong, Noh-Hee
    • Applied Chemistry for Engineering
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    • v.24 no.1
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    • pp.62-66
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    • 2013
  • This study was performed to investigate the physicochemical characteristics of glycoprotein extracted from carrot. We manufactured glycoprotein using natural vegetable raw materials carrots, and then analyzed it's physicochemical properties. The results showed that the nutritional composition of carrot glycoprotein (CG) consisted of 2.35% carbohydrate and 94.2% glycoprotein. There were small amounts of hydroxyproline and glycine known as characteristics of collagen peptide in the amino acid of CG, but there were large amount of glutamic acid and aspartic acid involved in the metabolism of glucose and fat. The calories of CG was 342.1 kcal/100 g and the molecular weight appeared less than 594 Da average molecular weight distribution.

Protective Effect of Glycoprotein Isolated from Cudrania tricuspidata on Liver in $CCl_4$-treated A/J Mice (생쥐에 있어서 꾸지뽕 당단백질의 간보호 효과)

  • Joo, Heon-Yeong;Lim, Kye-Taek
    • Korean Journal of Food Science and Technology
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    • v.41 no.1
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    • pp.93-99
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    • 2009
  • This study aimed to determine whether or not glycoprotein isolated from Cudrania tricuspidata Bureau fruit(CTB glycoprotein) exerts a hepatoprotective effect on liver injury induced by the administration of carbon tetrachloride($CCl_4$, 1.0mL/kg) to A/J mice. Following the administration of CTB glycoprotein(0-20mg/kg), the activities of antioxidant enzymes (superoxide dismutase(SOD), catalase(CAT), and glutathione peroxidase(GPx)), and the quantities of measured thiobarbituric acid reactive substances(TBARS), lactate dehydrogenase(LDH), and nitric oxide(NO) were evaluated from the murine liver tissues and plasma. Additionally, the activity of nuclear factor-kappa B(NF-${\kappa}B$) was assessed after pretreatment with $CCl_4$. When the mice were treated with $CCl_4$ alone, the activities of antioxidative enzymes reduced but amounts of TBARS, LDH, and NO increased. However, the results of treatment with CTB glycoprotein(10 and 20 mg/kg) revealed significantly increased activities of antioxidant enzymes(SOD, CAT, and GPx), as compared with $CCl_4$ alone. On the other hand, the result showed significant diminutions of the quantities of TBARS, LDH, and NO after treatment with CTB glycoprotein(10 and 20 mg/kg), as compared to $CCl_4$ alone. The activity of NF-${\kappa}B$ also declined after pretreatment with CTB glycoprotein, as compared with $CCl_4$ treatment alone. Thus, it is suggested that the CTB glycoprotein exerts a protective effect against $CCl_4$-induced liver injury in A/J mice.

Analysis of Non-segregated S-allele Strain by Single-Locus Hypothesis in Self-incompatible Brassica campestris (자가불화합성 Brassica campestris에 있어서 단일유전자좌가설에 의해 분리되지 않는 S-유전자 계통의 분석)

  • 노일섭
    • Journal of Plant Biology
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    • v.36 no.2
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    • pp.127-132
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    • 1993
  • Self-incompatibility in Brassica campestris is controlled by multi-allele system in a single genetic locus, the S locus, and it is elucidated that S-glycoproteins are S gene products. In this experiments, we examined the genetic mode(pollen tube behavior and segregation of S-glycoprotein), characteristic of S-glycoproteins and DNA constitution within nuclear genome on S gene family that unexplained by single locus model, and investigated the segregation pattern of S-glycoproteins in bred F1 generation. By diallel cross among the 15 plants within one family the existence of three types of homozygotes and three types of heterozygotes were observed, and segregation of S-allele could not explained by single locus model. From the results of IEF-immunoblot analysis for non-segregated individual plant, the segregation pattern of S specific bands was corresponded with results of diallel cross except with one case(SaSa genotype). The molecular weight of 6 different S-genotype varied in near by 50 kD, and each genotype expressed with 2 or 3 bands. Specific bands in SaSa, SbSb, ScSc has almost similar molecular weight between them. Southern analysis of genomic DNA probed with S-glycoprotein cDNA for 6 different genotypes revealed that there are clear difference in polymorphism, multiple bands of hybridization, when restriction enzymes of EcoR I were used. It could be assumed that there are several sequences related to the S-glycoprotein structural genes within their nuclear genome. Therefore, we suggested the possibilities that S-allele system could be controlled by multi-locus, that dominance-recessive interactions could be explained by modifier gene or supressor gene based on the results of abnormal segregation of S-glycoprotein in bred F1. The F2 analyses are progressing in now.

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Arginase-1 and P-glycoprotein are downregulated in canine hepatocellular carcinoma

  • Kim, Soo-Hyeon;Seung, Byung-Joon;Cho, Seung-Hee;Lim, Ha-Young;Bae, Min-Kyung;Sur, Jung-Hyang
    • Journal of Veterinary Science
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    • v.22 no.5
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    • pp.61.1-61.13
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    • 2021
  • Background: Hepatocellular carcinoma is the most common primary hepatic malignancy in humans and dogs. Several differentially expressed molecules have been studied and reported in human hepatocellular carcinoma and non-neoplastic liver lesions. However, studies on the features of canine hepatocellular carcinoma are limited, especially related to the differential characteristics of neoplastic and non-neoplastic lesions. Objectives: The study's objective was 1) to examine and evaluate the expression of arginase-1, P-glycoprotein, and cytokeratin 19 in canine liver tissues and 2) to investigate the differential features of hepatocellular carcinomas, liver tissue with non-neoplastic lesions, and paracancerous liver tissues in dogs. Methods: The expression levels of three markers underwent immunohistochemical analysis in 40 non-neoplastic liver tissues, 32 hepatocellular carcinoma tissues, and 11 paracancerous liver tissues. Scoring of each marker was performed semi-quantitatively. Results: Arginase-1 and P-glycoprotein were significantly downregulated in hepatocellular carcinoma, compared with hepatic tissues with non-neoplastic diseases (p < 0.001). Expression levels of arginase-1 and P-glycoprotein were also significantly lower in hepatocellular carcinoma than in paracancerous liver tissues (arginase-1, p = 0.0195; P-glycoprotein, p = 0.047). Few cytokeratin 19-positive hepatocytes were detected and only in one hepatocellular carcinoma and one cirrhotic liver sample. Conclusions: The results of this study suggest that downregulation of arginase-1 and P-glycoprotein is a feature of canine hepatocellular carcinoma; thus, those markers are potential candidates for use in differentiating hepatocellular carcinomas from non-neoplastic liver lesions in dogs.

Production of virus-like particles of nervous necrosis virus displaying partial VHSV's glycoprotein at surface and encapsulating DNA vaccine plasmids

  • Yang, Jeong In;Bessaid, Mariem;Kim, Ki Hong
    • Journal of fish pathology
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    • v.33 no.2
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    • pp.103-109
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    • 2020
  • In order to use nervous necrosis virus (NNV) virus-like particles (VLPs) as a delivery tool for heterologous antigens or plasmids, we attempted to produce red-spotted grouper nervous necrosis virus (RGNNV) VLPs displaying a partial region of viral hemorrhagic septicemia virus (VHSV) glycoprotein at the surface and VLPs that are harboring DNA vaccine plasmids within the VLP. A peptide encoding 105 amino acids of VHSV glycoprotein was genetically inserted in the loop region of NNV capsid gene, and VLPs expressing the partial part of VHSV glycoprotein were successfully produced. However, in the transmission electron microscope analysis, the shape and size of the partial VHSV glycoprotein-expressing NNV VLPs were irregular and variable, respectively, indicating that the normal assembly of capsid proteins was inhibited by the relatively long foreign peptide (105 aa) on the loop region. To encapsulate by simultaneous transformation with both NNV capsid gene expressing plasmids and DNA vaccine plasmids (having an eGFP expressing cassette under the CMV promoter), NNV VLPs containing plasmids were produced. The encapsulation of plasmids in the NNV VLPs was demonstrated by PCR and cells exposed to the VLPs encapsulating DNA vaccine plasmids showed fluorescence. These results suggest that the encapsulation of plasmids in NNV VLPs can be done with a simple one-step process, excluding the process of disassembly-reassembly of VLPs, and NNV VLPs can be used as a delivery tool for DNA vaccine vectors.

Analysis of the spike glycoprotein gene and nonstructural protein gene of transmissible gastroenteritis virus using PCR and RFLP analysis (PCR과 RFLP분석을 이용한 transmissible gastroenteritis virus의 spike glycoprotein gene과 nonstructural protein gene의 분석)

  • Kwon, Hyuk-moo
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.627-633
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    • 1996
  • To analyze the genomic diversity of transmissible gastroenteritis virus (TGEV), the N-terminal half of the spike (S) glycoprotein gene and nonstructural protein gene (open reading frames 3 and 3-1) were amplified by reverse transcriptase reaction and polymerase chain reaction (RT-PCR), and analyzed using restriction fragment length polymorphism (RFLP) patterns of the amplified DNA. In this study, TGEV Miller (M6) and Purdue (P115) strains were used as reference strains, and two vaccine strains (MSV and STC3) and four Korea isolates (P44, VRI-WP, VRI-41, and VRI-48) were analyzed. All TGEV strains were amplified with three TGEV primer pairs. Although there was some exception in RFLP analysis, this method differentiated TGEV strains into following groups : Miller group (M6 and MSV), Purdue group (PUS, STC3, P44, VRI-WP, VRI-41, and VRI-48). Using Sau3AI and SspI, VRI-48 was differentiated from the Miller and Purdue type viruses. The RT/PCR in conjuction with RFLP analysis was a rapid and valuable tool for differentiating several strains of TGEV. This study revealed the occurences of distinct difference in genome of TGEV strains.

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Hypolipidemic Effects of Glycoprotein Isolated from Ficus Carica Linnoeus in Mice (무화과 당단백질의 혈중지질 저하 효과)

  • Lim, Kye-Taek;Lee, Sei-Jung;Ko, Jeong-Hyeon;Oh, Phil-Sun
    • Korean Journal of Food Science and Technology
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    • v.37 no.4
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    • pp.624-630
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    • 2005
  • Glycoprotein (60 kDa) isolated from Ficus Carica Linnoeus (FCL glycoprotein) was examined by evaluating its hypolipidemic effects on plasma cholesterol levels and hepatic detoxicant enzyme activities in ICR mice. FCL glycoprotein $(100{\mu}g/mL)$ had strong scavenging activities (38%) against lipid peroxyl radicals. When mice were treated with Triton WR-1339 (400 mg/kg), levels of total cholesterol (TC) and low-density lipoprotein (LDL)-cholesterol in plasma significantly increased by 53.9 and 47.5 mg/dL, respectively, compared to the control, whereas, when pretreated with FCL glycoprotein $(100{\mu}g/mL)$, decreased remarkably by 55.4, and 47,0 mg/dL, compared to Triton WR-1339 treatment alone. Interestingly, high-density lipoprotein (HDL)-cholesterol level did not change. Body and liver weights did not change significantly after Triton WR-1339 treatment in presence of FCL glycoprotein. FCL glycoprotein $(100{\mu}g/mL)$ stimulated activities of antioxidative detoxicant enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), whereas GPx activity significantly increased compared to the control. These results suggest FCL glycoprotein has abilities to scavenge lipid peroxyl radicals, lower plasma lipid levels, and stimulate detoxicant enzyme activity in mouse liver.