• 제목/요약/키워드: viral protein 2

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토마토에서 분리된 담배 모자이크 바이러스 외피단백질 유전자의 cDNA 클로닝 및 염기서열 분석 (Complementary DNA Cloning and Nucleotide Sequence Analysis of Coat Protein Gene from TMV Tomato Strain)

  • 이청호;이영기;강신웅;박은경
    • 한국연초학회지
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    • 제18권2호
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    • pp.101-106
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    • 1996
  • Tobacco mosaic virus (TMV) tomato strain was isolated from tomato "Seo-Kwang" in Korea. The virion was purified by density gradient centrifugation, and total viral RNA was isolated from the purified particles. Coat protein (CP) cDNA of the virus was synthesized by RT-PCR, and the purified cDNA fragment was subcloned to pBluescript II SK-. The analysis of nucleotide sequence showed that this cDNA was 693 nucleotides long from the insert of clone p1571 and p1572 which contain complete codons of the viral coat protein gene (474 nucleotides) and 3' untranslated region. The nucleotides of coat protein encoding cDNA of the strain were 6 nucleotides less than that of TMV common strain isolated from tobacco plant in Korea. The CP gene showed 70% maximum homology with that of the common strain in the nucleotide level and 86% maximum homology in amino acid level.cid level.

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Comparison of COVID-19 Vaccines Introduced in Korea

  • Lee, Chang-Gun;Lee, Dongsup
    • 대한의생명과학회지
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    • 제28권2호
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    • pp.67-82
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    • 2022
  • The prevalence of SARS-CoV-2 led to inconsistent public health policies that resulted in COVID-19 containment failure. These factors resulted in increased hospitalization and death. To prevent viral spread and achieve herd immunity, the only safe and effective measure is to provide to vaccinates. Ever since the release of the SARS-CoV-2 nucleotide sequence in January of 2020, research centers and pharmaceutical companies from many countries have developed different types of vaccines including mRNA, recombinant protein, and viral vector vaccines. Prior to initiating vaccinations, phase 3 clinical trials are necessary. However, no vaccine has yet to complete a phase 3 clinical trial. Many products obtained "emergency use authorization" from governmental agencies such as WHO, FDA etc. The Korean government authorized the use of five different vaccines. The viral vector vaccine of Oxford/AstraZeneca and the Janssen showed effectiveness of 76% and 66.9%, respectively. The mRNA vaccine of Pfizer-BioNTech and Moderna showed effectiveness of 95% and 94.1%, respectively. The protein recombinant vaccine of Novavax showed an effectiveness of 90.4%. In this review, we compared the characteristics, production platform, synthesis principles, authorization, protective effects, immune responses, clinical trials and adverse effects of five different vaccines currently used in Korea. Through this review, we conceptualize the importance of selecting the optimal vaccine to prevent the COVID-19 pandemic.

Infectious RNA Viruses in the Edible Mushroom Pleurotus spp.

  • Park, Jeonga-Soo;Kim, Young-Ho
    • Journal of Microbiology
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    • 제34권1호
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    • pp.61-67
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    • 1996
  • Double-stranded RNA (dsRNA) viruses and single-stranded RNA(ssRNA) viruses were detected in a strain of Pleurotus mushroom cultivated in a farm. Those fungal virsus were purified in the pH 6.0 or pH 7.2 using CsCI or Cs$_{2}$SO$_{4}$ buoyant density centrifugation. Each viral particles were not completely separated at any trials. However, mushroom bacili-form virus contains a single major nucleic acid with 0.7 Kb ssRNA, which might code for 20 Kd viral capsid protein. The dsRNAs are encapsidatred into spherical-form viruses, whereas ssRNA viral genomes are encapsidated into two different sizes of bacili-form particles. A healthy-looking mushroom also contained some spherical-form viruses with dsRNAs. Laboratory strains of Pleurotus ostreatus and a cultivated strain of P. sajor-caju did not show any viral particles. Mushrooms with specific disease symptoms. however, contained at least four different sizes of spherical-form viruses. Thus, we concluded that a bacilli-form virus case a severe disease symptoms of adnormal on mushroom development.

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Identification of Grapevine leafroll-associated virus 3 Ampelovirus from Grapevines in Korea

  • Kim, Hyun-Ran;Lee, Sin-Ho;Lee, Bong-Choon;Kim, Yeong-Tae;Park, Jin-Woo
    • The Plant Pathology Journal
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    • 제20권2호
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    • pp.127-130
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    • 2004
  • Grapevine leaf roll-associated virus 3 (GLRaV-3) is one of the most important viral diseases of grapevine in the world. In this study, GLRaV-3 Ampelovirus was identi-fied from grapevines in Korea by analyzing viral coat protein size, nucleotide, and amino acid sequences. The molecular weight of viral coat protein from virus-infected in vitro plantlets was determined by western blot using a commercial GLRaV-3 polyclonal antibody. Western blot analysis showed a coat protein of about 43 kDa. RT-PCR product of about 942 bp which encoded the coat protein (CP) gene was amplified with specific primers. When the viruses existed at low titers in the host plant, the dsRNA had very specific template in RT- PCR amplification of fruit tree viruses. Especially, small-scale dsRNA extraction method was very reliable and rapid. Sequence analysis revealed that the CP of the GLRaV-3 Ko consisted of 942 bp nucleotide, which encoded 314 amino acid residues. The CP gene of GLRaV-3 Ko had 98.9% nucleotide sequence and 98.7% amino acid sequence identities with earlier reported GLRaV-3. This is the first report on molecular assay of GLRaV-3 Ampelovirus identified from Korea. The GLRaV-3 Ko CP clone would be very useful for breeding of virus resistant grapevines.

Mutational Analysis of an Essential RNA Stem-loop Structure in a Minimal RNA Substrate Specifically Cleaved by Leishmania RNA Virus 1-4 (LRV1-4) Capsid Endoribonuclease

  • Ro, Youngtae;Patterson, Jean L.
    • Journal of Microbiology
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    • 제41권3호
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    • pp.239-247
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    • 2003
  • The LRV1-4 capsid protein possesses an endoribonuclease activity that is responsible for the single site-specific cleavage in the 5' untranslated region (UTR) of its own viral RNA genome and the formation of a conserved stem-loop structure (stem-loop IV) in the UTR is essential for the accurate RNA cleavage by the capsid protein. To delineate the nucleotide sequences, which are essential for the correct formation of the stem-loop structure for the accurate RNA cleavage by the viral capsid protein, a wildtype minimal RNA transcript (RNA 5' 249-342) and several synthetic RNA transcripts encoding point-mutations in the stem-loop region were generated in an in vitro transcription system, and used as substrates for the RNA cleavage assay and RNase mapping studies. When the RNA 5' 249-342 transcript was subjected to RNase T1 and A mapping studies, the results showed that the predicted RNA secondary structure in the stem-loop region using FOLD analysis only existed in the presence of Mg$\^$2+/ ions, suggesting that the metal ion stabilizes the stem-loop structure of the substrate RNA in solution. When point-mutated RNA substrates were used in the RNA cleavage assay and RNase T1 mapping study, the specific nucleotide sequences in the stem-loop region were not required for the accurate RNA cleavage by the viral capsid protein, but the formation of a stem-loop like structure in a region (nucleotides from 267 to 287) stabilized by Mg$\^$2+/ ions was critical for the accurate RNA cleavage. The RNase T1 mapping and EMSA studies revealed that the Ca$\^$2+/ and Mn$\^$2+/ ions, among the reagents tested, could change the mobility of the substrate RNA 5' 249-342 on a gel similarly to that of Mg$\^$2+/ ions, but only Ca$\^$2+/ ions identically showed the stabilizing effect of Mg$\^$2+/ ions on the stem-loop structure, suggesting that binding of the metal ions (Mg$\^$2+/ or Ca$\^$2+/) onto the RNA substrate in solution causes change and stabilization of the RNA stem-loop structure, and only the substrate RNA with a rigid stem-loop structure in the essential region can be accurately cleaved by the LRV1-4 viral capsid protein.

Modulation of a Fungal Signaling by Hypovirus

  • Kim, Dae-Hyuk
    • The Plant Pathology Journal
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    • 제19권1호
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    • pp.30-33
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    • 2003
  • The chestnut blight fungus, Cryphonectria parasitica, and its hypovirus aye a useful model system in the study of the mechanisms of hypoviral infection and its consequences, such as a biological control of fungal pathogens. Strains containing the double-stranded (ds) RNA viruses Cryphonectria hypovirus 1 show characteristic symptoms of hypovirulence and display hypovirulence-associated changes, such as reduced pigmentation, sporulation, laccase production, and oxalate accumulation. Interestingly, symptoms caused by hypoviral infection appear to be the result of aberrant expression of a number of specific genes in the hypovirulent strain. Several viral regulated fungal genes are identified as cutinase gene, Lac1, which encodes an extracellular laccase, Crp, which encodes an abundant tissue-specific cell-surface hydrophobin that mediates physical strength, and Mf2/1 and Mf2/2, which encode pheromone genes involved in poor sporulation in the presence of hypo-virus. Since the phenotypic changes in the fungal host are pleiotropic, although coordinated and specific, it has been suggested that the hypovirus disturbs one or several regulatory pathways (Nuss,1996). Accordingly, several studies have shown the implementation of a signal transduction pathway during viral symptom development. Although further studies are required, hypovirulence and its associated symptom development due to the hypoviral regulation of a fungal hetero-trimeric G-protein have been suggested. In addition, recent studies have shown the presence of a novel protein kinase gene cppk1 and its transcriptional upregulation by hypovirus. In this review, the presence of important components in signal transduction pathway, their putative biological function, and viral-specific regulation will be addressed.

Distinct Humoral and Cellular Immunity Induced by Alternating Prime-boost Vaccination Using Plasmid DNA and Live Viral Vector Vaccines Expressing the E Protein of Dengue Virus Type 2

  • George, Junu A.;Eo, Seong-Kug
    • IMMUNE NETWORK
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    • 제11권5호
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    • pp.268-280
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    • 2011
  • Background: Dengue virus, which belongs to the Flavivirus genus of the Flaviviridae family, causes fatal dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) with infection risk of 2.5 billion people worldwide. However, approved vaccines are still not available. Here, we explored the immune responses induced by alternating prime-boost vaccination using DNA vaccine, adenovirus, and vaccinia virus expressing E protein of dengue virus type 2 (DenV2). Methods: Following immunization with DNA vaccine (pDE), adenovirus (rAd-E), and/or vaccinia virus (VV-E) expressing E protein, E protein-specific IgG and its isotypes were determined by conventional ELISA. Intracellular CD154 and cytokine staining was used for enumerating CD4+ T cells specific for E protein. E protein-specific CD8+ T cell responses were evaluated by in vivo CTL killing activity and intracellular IFN-${\gamma}$ staining. Results: Among three constructs, VV-E induced the most potent IgG responses, Th1-type cytokine production by stimulated CD4+ T cells, and the CD8+ T cell response. Furthermore, when the three constructs were used for alternating prime-boost vaccination, the results revealed a different pattern of CD4+ and CD8+ T cell responses. i) Priming with VV-E induced higher E-specific IgG level but it was decreased rapidly. ii) Strong CD8+ T cell responses specific for E protein were induced when VV-E was used for the priming step, and such CD8+ T cell responses were significantly boosted with pDE. iii) Priming with rAd-E induced stronger CD4+ T cell responses which subsequently boosted with pDE to a greater extent than VV-E and rAd-E. Conclusion: These results indicate that priming with live viral vector vaccines could induce different patterns of E protein-specific CD4+ and CD8+ T cell responses which were significantly enhanced by booster vaccination with the DNA vaccine. Therefore, our observation will provide valuable information for the establishment of optimal prime-boost vaccination against DenV.

Acibenzolar-S-Methyl(ASM)-Induced Resistance against Tobamoviruses Involves Induction of RNA-Dependent RNA Polymerase(RdRp) and Alternative Oxidase(AOX) Genes

  • Madhusudhan, Kallahally Nagendra;Deepak, Saligrama Adavigowda;Prakash, Harishchandra Sripathi;Agrawal, Ganesh Kumar;Jwa, Nam-Soo;Rakwal, Randeep
    • Journal of Crop Science and Biotechnology
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    • 제11권2호
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    • pp.127-134
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    • 2008
  • Tobamoviruses are the major viral pathogens of tomato and bell pepper. The preliminary results showed that Acibenzolar-Smethyl(ASM; S-methylbenzo(1,2,3) thiadiazole-7-carbothiate) pre-treatment to tomato and tobacco plants reduces the concentration of Tomato mosaic tobamovirus(ToMV) and Tobacco mosaic tobamovirus(TMV) in tomato and bell pepper seedlings, respectively. Pre-treatment of the indicator plant(Nicotiana glutinosa) with the ASM followed by challenge inoculation with tobamoviruses produced a reduced number and size of local lesions(67 and 79% protection over control to TMV and ToMV inoculation, respectively). In order to understand the mechanism of resistance the gene expression profiles of antiviral genes was examined. RT-PCR products showed higher expression of two viral resistance genes viz., alternative oxidase(AOX) and RNA dependent RNA polymerase(RdRp) in the upper leaves of the ASM-treated tomato plants challenge inoculation with ToMV. Further, the viral concentration was also quantified in the upper leaves by reverse transcription PCR using specific primer for movement protein of ToMV, as well as ELISA by using antisera against tobamoviruses. The results provided additional evidence that ASM pre-treatment reduced the viral movement to upper leaves. The results suggest that expressions of viral resistance genes in the host are the key component in the resistance against ToMV in the inducer-treated tomato plants.

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Molecular detection of bat coronaviruses in three bat species in Indonesia

  • Dharmayanti, Ni Luh Putu Indi;Nurjanah, Diana;Nuradji, Harimurti;Maryanto, Ibnu;Exploitasia, Indra;Indriani, Risa
    • Journal of Veterinary Science
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    • 제22권6호
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    • pp.70.1-70.12
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    • 2021
  • Bats are an important reservoir of several zoonotic diseases. However, the circulation of bat coronaviruses (BatCoV) in live animal markets in Indonesia has not been reported. Genetic characterization of BatCoV was performed by sequencing partial RdRp genes. Real-time polymerase chain reaction based on nucleocapsid protein (N) gene and Enzyme-linked immunosorbent assay against the N protein were conducted to detect the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral RNA and antibody, respectively. We identified the presence of BatCoV on Cynopterus brachyotis, Macroglossus minimus, and Rousettus amplexicaudatus. The results showed that the BatCoV included in this study are from an unclassified coronavirus group. Notably, SARS-CoV-2 viral RNA and antibodies were not detected in the sampled bats.