• Title/Summary/Keyword: viral/genetics

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Complete Genome Sequences of the Genomic RNA of Soybean mosaic virus Strains G7B and G5

  • Kim, Kook-Hyung;Lim, Won-Seok;Kim, Yul-Ho
    • The Plant Pathology Journal
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    • v.19 no.3
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    • pp.171-176
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    • 2003
  • The complete nucleotide sequences of the genomic RNAs of Soybean mosaic virus strains GS (SMV-G5) and G7H (SMV-G7H) were determined and compared with sequences of other SMV strains. Each viral RNA was determined to be 9588 nucleotides in length excluding the poly (A) tail and contained an open reading frame to encode a polyprotein subsequently processed into up to ten proteins by proteolytic cleavage. Com-parison of the amino acid sequences with those of other SMV strains showed high percentage of amino acid sequence homology with the same genome organization. The nucleotide and the deduced amino acid sequences between SMV-G5 and SMV-G7H were greater than 99% identity. When compared with those of other SMV strains in a phylogenetic analysis of the nucleotide and deduced amino acid sequences, they formed a distinct virus clade showing over 97% amino acid identity, but were more distantly related to the other potyvirus (44.1-69.6% identity). Interestingly, SMV G7H strain caused a severe mosaic or necrosis symptom in soybean cultivars including Jinpum-1, Jinpum-2, and Sodam, whereas, no symptom was observed in SMV-G5 inoculation. Complete nucleotide sequences of these strains will give clues for determining symptom determinant(s) in future research.

COVID-19: an update on diagnostic and therapeutic approaches

  • Iyer, Mahalaxmi;Jayaramayya, Kaavya;Subramaniam, Mohana Devi;Lee, Soo Bin;Dayem, Ahmed Abdal;Cho, Ssang-Goo;Vellingiri, Balachandar
    • BMB Reports
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    • v.53 no.4
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    • pp.191-205
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    • 2020
  • The unexpected pandemic set off by the novel coronavirus 2019 (COVID-19) has caused severe panic among people worldwide. COVID-19 has created havoc, and scientists and physicians are urged to test the efficiency and safety of drugs used to treat this disease. In such a pandemic situation, various steps have been taken by the government to control and prevent the Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2). This pandemic situation has forced scientists to rework strategies to combat infectious diseases through drugs, treatment, and control measures. COVID-19 treatment requires both limiting viral multiplication and neutralizing tissue damage induced by an inappropriate immune reaction. Currently, various diagnostic kits to test for COVID-19 are available, and repurposing therapeutics for COVID-19 has shown to be clinically effective. As the global demand for diagnostics and therapeutics continues to rise, it is essential to rapidly develop various algorithms to successfully identify and contain the virus. This review discusses the updates on specimens/samples, recent efficient diagnostics, and therapeutic approaches to control the disease and repurposed drugs mainly focusing on chloroquine/hydroxychloroquine and convalescent plasma (CP). More research is required for further understanding of the influence of diagnostics and therapeutic approaches to develop vaccines and drugs for COVID-19.

Virulence Differentiation of Eight Turnip mosaic virus Isolates Infecting Cruciferous Crops

  • Choi, Hong-Soo;Sohn, Seong-Han;Yoon, Moo-Kyoung;Cheon, Jeong-Uk;Kim, Jeong-Soo;Were, Hassan Karakacha;Cho, Jang-Kyung;Kim, Kook-Hyung;Takanami, Yoichi
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.369-376
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    • 2005
  • Turnip mosaic virus (TuMV) is an infectious viral pathogen on the cruciferous crops, predominantly Chinese cabbage (Brassica campestris subsp. pekinensis) and radish (Raphanus sativus). On the basis of the symptom development in selective differential hosts from indicator host species, Chinese cabbage and Korean radish inbred lines, the representative eight isolates of TuMV were divided into two major groups/or six types. Group I includes Th 1, Ca-ad7, and Cj-ca2-1 isolates, while group II includes the other isolates (rg-pfl, r 9-10, Rhcql-2, Stock and Mustard). According to the molecular phylogenetic analysis, these isolates, however, divided into two groups and two independent isolates. Phylogenetic analysis indicated that four isolates (Tu 1, r9-10, Stock and Rh-cql-2) formed a distinct phylogenetic group, and the other two isolates (Ca-ad7 and Cj-ca2-1) also formed another group. Mustard and rg-pfl isolates did not seem to have any relationship with these two groups. Taken together, these results indicated that virulence differentiation on host plants, molecular phylogenetic analysis of the nucleotide and the deduced amino acid of TuMV coat proteins did not show any relationship. The multi-resistant lines, Wonyae 20026 and BP058 in Chinese cabbage represent valuable genetic materials that can be used for crucifer breeding programs on TuMV resistance, but not in Korean radish.

The SL1 Stem-Loop Structure at the 5′-End of Potato virus X RNA Is Required for Efficient Binding to Host Proteins and forViral Infectivity

  • Kwon, Sun-Jung;Kim, Kook-Hyung
    • Molecules and Cells
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    • v.21 no.1
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    • pp.63-75
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    • 2006
  • The 5′-region of Potato virus X (PVX) RNA, which contains an AC-rich, single-stranded region and stem-loop structure 1 (SL1), affects RNA replication and assembly. Using Systemic Evolution of Ligands by EXponential enrichment (SELEX) and the electrophoretic mobility shift assay, we demonstrate that SL1 interacts specifically with tobacco protoplast protein extracts (S100). The 36 nucleotides that correspond to the top region of SL1, which comprises stem C, loop C, stem D, and the tetra loop (TL), were randomized and bound to the S100. Remarkably, the wild-type (wt) sequence was selected in the second round, and the number of wt sequences increased as selection proceeded. All of the selected clones from the fifth round contained the wt sequence. Secondary structure predictions (mFOLD) of the recovered sequences revealed relatively stable stem-loop structures that resembled SL1, although the nucleotide sequences therein were different. Moreover, many of the clones selected in the fourth round conserved the TL and C-C mismatch, which suggests the importance of these elements in host protein binding. The SELEX clone that closely resembled the wt SL1 structure with the TL and C-C mismatch was able to replicate and cause systemic symptoms in plants, while most of the other winners replicated poorly only on inoculated leaves. The RNA replication level on protoplasts was also similarly affected. Taken together, these results indicate that the SL1 of PVX interacts with host protein(s) that play important roles related to virus replication.

Application of Antimicrobial Peptide LL-37 as an Adjuvant for Middle East Respiratory Syndrome-Coronavirus Antigen Induces an Efficient Protective Immune Response Against Viral Infection After Intranasal Immunization

  • Ju Kim;Ye Lin Yang;Yongsu Jeong;Yong-Suk Jang
    • IMMUNE NETWORK
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    • v.22 no.5
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    • pp.41.1-41.16
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    • 2022
  • The human antimicrobial peptide LL-37 has chemotactic and modulatory activities in various immune cells, including dendritic cells. Because of its characteristics, LL-37 can be considered an adjuvant for vaccine development. In this study, we confirmed the possible adjuvant activity of LL-37 in mucosal vaccine development against Middle East respiratory syndrome-coronavirus (MERS-CoV) by means of intranasal immunization in C57BL/6 and human dipeptidyl peptidase 4 (hDPP4)-transgenic (hDPP4-Tg) mice. Intranasal immunization using the receptor-binding domain (RBD) of MERS-CoV spike protein (S-RBD) recombined with LL-37 (S-RBD-LL-37) induced an efficient mucosal IgA and systemic IgG response with virus-neutralizing activity, compared with S-RBD. Ag-specific CTL stimulation was also efficiently induced in the lungs of mice that had been intranasally immunized with S-RBD-LL-37, compared with S-RBD. Importantly, intranasal immunization of hDPP4-Tg mice with S-RBD-LL-37 led to reduced immune cell infiltration into the lungs after infection with MERS-CoV. Finally, intranasal immunization of hDPP4-Tg mice with S-RBD-LL-37 led to enhanced protective efficacy, with increased survival and reduced body weight loss after challenge infection with MERS-CoV. Collectively, these results suggest that S-RBD-LL-37 is an effective intranasal vaccine candidate molecule against MERS-CoV infection.

A Molecular Study of Rice Black-Streaked Dwarf Virus (벼 흑조위축병 바이러스의 분자생물학적 연구)

  • Park, Jong-Sug;Bae, Shin-Chyul;Kim, Young-Min;Paik, Young-Ki;Kim, Ju-Kon;Hwang, Young-Soo
    • Applied Biological Chemistry
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    • v.37 no.3
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    • pp.148-153
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    • 1994
  • Rice black-streaked dwarf virus (RBSDV), a member of the plant reoviridae fijivirus group, causes a serious damage for rice production in Korea. To characterize the RBSDV genome, virus particles were produced by feeding of planthopper (Laodelphax striatellus F.) carring RBSDV to maize plants for 2 days. In $30{\sim}40$ days after feeding, the viral particles were purified from the infected maize roots by using $10{\sim}40%$ sucrose gradient centrifugation. After treatment of 10% SDS to remove the viral coat proteins, ten viral double-stranded RNAs were resolved in agrose gel electrophoresis. Total dsRNA was then used to synthesize cDNA by reverse transcriptase and a cDNA library was constructed in the ${\lambda}gt11$ vector. The phages that contain RBSDV cDNA fragments were selected by hybridizing with the random-primed probe prepared from RBSDV dsRNAs. After subcloning of several cDNA fragments into the pUC19 plasmid vector, one clone (pRV3) was chosen for sequencing. The pRV3 clone was shown to be located on the RBSDV genome fragment No.3 by RNA gel-blot analysis. Sequence analysis of the clone revealed that the pRV3 contains two partial open reading frames.

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Induction of RNA-mediated Resistance to Papaya Ringspot Virus Type W

  • Krubphachaya, Pongrit;Juricek, Mila;Kertbundit, Sunee
    • BMB Reports
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    • v.40 no.3
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    • pp.404-411
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    • 2007
  • Transformation of cantaloupes with the coat protein (cp) gene of papaya ringspot virus type W (PRSV-W), Thai isolate, was used to introduce virus resistance. Binary vectors containing either the full length coat protein coding region under control of the 35S CaMV promoter(pSA1175), or the inverted-repeat of a coat protein coding region (pSA1304), were constructed and used for Agrobacteriummediated transformation of cotyledonary explants of the cantaloupe cultivar Sun Lady. Four independent transgenic lines were obtained using pSA1304 and one using pSA1175. Integration of the PRSV-W cp gene into the genome of these transgenic lines was verified by PCR amplification, GUS assays and Southern blot hybridization. In vitro inoculation of these lines with PRSV-W revealed that whereas the line containing pSA1175 remained sensitive, the four lines containing pSA1304 were resistant. The presence of small RNA species, presumably siRNA, corresponding to regions of the viral cp gene in transgenic lines resistant to PRSV-W supports the involvement of post-transcriptional gene silencing in the establishment of resistance.

Growth and Yield Comparisons among Barley Varieties with Different Resistance to Barley Mosaic Virus (보리 모자이크 바이러스 저항성 정도가 상이한 보리 품종간 생육 및 수량 비교)

  • Park Jong-Chul;Lee Mi-Ja;Choi In-Bae;Kim Mi-Jung;Park Chul-Soo;Kim Jung-Gon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.5
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    • pp.477-482
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    • 2006
  • Viral diseases, especially Barley yellow mosaic virus (BaYMV) and Barley mud mosaic virus (BaMMV), have been most serious in barley fields. In this study, we investigated the effect of different level of resistance to viral diseases on the plant growth and yield in barley. Various viral disease symptoms on leaves of overwintered plants were similar between medial-resistant and susceptible varieties of Saessalbori and Baegdong. In diagnosis of virus infection, BaYMV and BaMMV were detected in Saessalbori and Baegdong, but not in the resistant variety, Naehanssalbori. Plant height was restrained about $11{\sim}12cm$ prior to heading in Saessalbori and Baegdong comparing to Naehanssalbori. Even if both varieties were medial resistant to virus diseases, Saessalbori was different from Baegdong in heading date and culm length due to its recovery from viral damages prior to heading. Both medial-resistant and susceptible varieties were quite different from the resistant variety in yield components such as heading date, number of spikes and culm length when evaluated in the virus-infected or non-infected field. Baegdong delayed 7 days in heading date and reduced by more than 50% in culm length and spike numbers as compared to Naehanssalbori. On the other hand, Saessalbori showed similar heading date, but was shorter by 20% in culm length than Naehanssalbori. Three varieties tested in the non-infected field over two years were not significantly different for yield potential with ranges of $340{\sim}405kg/10a$. However, significant yield reduction (P<0.01) was observed in Saessalbori and Baegdong with ranges of $108{\sim}288kg/10a$ as compared to Naehanssalbori (391 kg/10a) when tested in the virus-infected field. Yield potentials of Saessalbori and Baegdong reduced by 35 and 63%, respectively in the virus-infected field as compared to those in the non-infected field. Our results showed that damages from virus diseases were significant on the early plant growth to yield and its components in barley.

Double Mutations in eIF4E and eIFiso4E Confer Recessive Resistance to Chilli Veinal Mottle Virus in Pepper

  • Hwang, JeeNa;Li, Jinjie;Liu, Wing-Yee;An, Song-Ji;Cho, Hwajin;Her, Nam Han;Yeam, Inhwa;Kim, Dosun;Kang, Byoung-Cheorl
    • Molecules and Cells
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    • v.27 no.3
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    • pp.329-336
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    • 2009
  • To evaluate the involvement of translation initiation factors eIF4E and eIFiso4E in Chilli veinal mottle virus (ChiVMV) infection in pepper, we conducted a genetic analysis using a segregating population derived from a cross between Capsicum annuum 'Dempsey' containing an elF4E mutation ($pvr1^2$) and C. annuum 'Perennial' containing an elFiso4E mutation (pvr6). C. annuum 'Dempsey' was susceptible and C. annuum 'Perennial' was resistant to ChiVMV. All $F_1$ plants showed resistance, and $F_2$ individuals segregated in a resistant-susceptible ratio of 166:21, indicating that many resistance loci were involved. Seventy-five $F_2$ and 329 $F_3$ plants of 17 families were genotyped with $pvr1^2$ and pvr6 allele-specific markers, and the genotype data were compared with observed resistance to viral infection. All plants containing homozygous genotypes of both $pvr1^2$ and pvr6 were resistant to ChiVMV, demonstrating that simultaneous mutations in elF4E and eIFiso4E confer resistance to ChiVMV in pepper. Genotype analysis of $F_2$ plants revealed that all plants containing homozygous genotypes of both $pvr1^2$ and pvr6 showed resistance to ChiVMV. In protein-protein interaction experiments, ChiVMV viral genome-linked protein (VPg) interacted with both eIF4E and eIFiso4E. Silencing of elF4E and eIFiso4E in the VIGS experiment showed reduction in ChiVMV accumulation. These results demonstrated that ChiVMV can use both eIF4E and eIFiso4E for replication, making simultaneous mutations in eIF4E and eIFiso4E necessary to prevent ChiVMV infection in pepper.

Effective Antitumor Activity of a Recombinant Vaccinia Virus Expressing Murine Interleukin 4 (인터루킨-4를 발현하는 재조합 백시니아 바이러스에 의한 암성장의 억제)

  • Yoon, Kee-Jung;Jin, Ning-Yi;Kim, Sun-Young
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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    • pp.71-78
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    • 1998
  • Vaccinia virus is the prototype orthopoxvirus that has been used as a vaccine strain for small pox. This virus has been used to express a variety of cellular and viral genes in mammalian cells at high levels. Interleukin-4 (IL-4) has been found to stimulate the proliferation of T cells and enhance the cytolytic activity of cytotoxic T lymphocytes. To test the immunotherapeutic potential of IL-4 delivered in vivo by poxvirus, a recombinant vaccinia virus expressing the murine IL-4 gene (RVVmIL-4) was constructed. A high level of IL-4 production was confirmed by infecting HeLa cells and measuring IL-4 in cell culture supernatant by ELISA. As a tumor model, two cell lines were used; the murine T leukemic line P388 and the murine breast cancer line TS/A. CDF1 mice were intraperitoneally inoculated with $1\;{\times}\;10^5$ cells of P388. Mice were injected at the same site with $5\;{\times}\;10^5\;PFU$ of recombinant vaccinia virus; first, 3 days after the injection of tumor cells and thereafter once every week for 3 weeks. Intraperitoneal injections of RVVmIL-4 significantly prolonged the survival time of mice inoculated with tumor cells. All mice injected with RVVmIL-4 remained alive for 30 days after the postinoculation of tumor cells, while 100% and 70% of the animals injected with saline or wild type vaccinia virus died, respectively. In another tumor model using TS/A, tumor was established by subcutaneously inoculating $2{\times}10^5$ tumor cells to BALB/c mice. After tumor formation was confirmed on day 4 in all mice, $5\;{\times}\;10^6\;PFU$ of RVVmIL-4 was inoculated subcutaneously three times, once every week for 3 weeks. The TS/A tumor was eradicated in two of the nine mice. Seven of the nine mice treated with RVVmIL-4 developed a tumor, but tumor growth was significantly delayed compared to those treated with saline or wild type vaccinia virus. These results indicate that recombinant vaccinia viruses may be used as a convenient tool for delivering immunomodulator genes to a variety of tumors.

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