• 제목/요약/키워드: Ds-isolates

검색결과 38건 처리시간 0.023초

Cloning and Purification of Envelope Proteins (VP19, VP28) and Nucleocapsid Proteins (VP15, VP35) Genes of a Shrimp White Spot Syndrome Virus Isolates in Korea

  • Seok, Seung-hyeok;Park, Jae-hak
    • 한국수의병리학회:학술대회논문집
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    • 한국수의병리학회 2003년도 추계학술대회초록집
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    • pp.41-41
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    • 2003
  • White spot syndrome virus (WSSV) is the causative agent of a disease that has led to severe mortalities of cultured shrimps in Korea and many other countries. Since 1993, massive mortalities due to the viral infection have also occurred in the penaeid shrimps cultured in Korea. WSSV is a large, circular, double stranded (ds) DNA virus and an enveloped, ellipsoid virus with a rod-shaped nucleocapsid with flat ends. In order to identify the characteristics of this Korean isolate of WSSV, the genes for four virion proteins, VP15, VP19, VP28 and VP35 were cloned and their sequences were compared with the available pool of WSSV gene sequences in the GenBank/EMBL databases. From these comparisons, we confirm the occurrence of WSSV in Korea and deduce that, VP15, VP28 and VP35 genes are identically conserved among the Korean isolate and geographically different foreign isolates, but VP19 amino acid sequences of the Korean WSSV isolates changed valine of the foreign isolates into aspartate. (omitted)

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Identification of a New Potyvirus Associated with Chlorotic Vein Banding Disease of Spathiphyllum spp., in Andhra Pradesh, India

  • Padmavathi, M.;Srinivas, K.P.;Reddy, Ch. V. Subba;Ramesh, B.;Navodayam, K.;Krishnaprasadji, J.;Babu, P. Ratan;Sreenivasulu, P.
    • The Plant Pathology Journal
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    • 제27권1호
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    • pp.33-36
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    • 2011
  • The genome of a potyvirus isolate associated with chlorotic spots and vein banding symptoms on Spathiphyllum spp., in Andhra Pradesh state, India was amplified by RT-PCR using degenerate potyvirus primers, amplicons cloned, and sequence (1.6 kb) analyzed. This virus isolate shared maximum identity of 74.8% and 80.2% at coat protein (CP) gene nucleotide (906 nucleotides) and amino acid (302 amino acids) levels, respectively with Dasheen mosaic virus (DsMV)-M13 isolate reported from China. But its 3'-UTR (258 nucleotides) had maximum identity of 62.5% with DsMV-Vietnam isolate. The deduced molecular weight of CP is 33.57 kDa and it contained DAG triplet in its N-terminal region. In CP amino acid based phylogenetic analysis, this virus isolate represented a separate branch but closer to DsMV isolates cluster. Based on the molecular criteria set for the discrimination of species and genus in the Potyviridae family, the present virus isolate was identified as a distinct virus species in the genus Potyvirus and proposed the name Spathiphyllum chlorotic vein banding virus (SCVbV).

Biological Control of Some Serious Weeds in Dakahlia District. II. Mycoherbicial Production and Physiological Host Responses

  • Abdel-Fattah, Gamal M.
    • Mycobiology
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    • 제30권2호
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    • pp.96-101
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    • 2002
  • Four pathogenic fungal isolates belonging to different genera including Alternaria, Fusarium and Curvularia were isolated from selected diseased weeds growing in the fields in Dakahalia district. The inoculum of these pathogenic fungi specific to weeds were cultured, standardized and formulated as alginate pellets containing mycelium plus culture filtrate. These mycoherbicides were evaluated for disease severity(DS). Maximum DS was obtained with the alginate pellets of mycelium filtrate Fusarium solani. Physiological changes of the treated weed were determined 5 aiid 10 days after treatments. As compared to the healthy weeds, all mycoherbicide formulations significantly decreased the amount of photosynthetic pigments and subsequently soluble and insoluble sugars in the infected weeds. The mycoherbicide formulation of F. solani had the greatest effect on lowering to the abovementioned amount in the leaves of Chenopodium murale. Generally, treatment of weed leaves with the specific mycoherbicide led to a highly significant increase in total phenol content when compared to the healthy control weed. C. murale infected with the mycoherbicide formulation of F. solani had higher levels of phenolic compounds than those other treated weeds particularly after 10 days of inoculation.

Differential Protein Expressions in Virus-Infected and Uninfected Trichomonas vaginalis

  • Ding, He;Gong, Pengtao;Yang, Ju;Li, Jianhua;Li, He;Zhang, Guocai;Zhang, Xichen
    • Parasites, Hosts and Diseases
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    • 제55권2호
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    • pp.121-128
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    • 2017
  • Protozoan viruses may influence the function and pathogenicity of the protozoa. Trichomonas vaginalis is a parasitic protozoan that could contain a double stranded RNA (dsRNA) virus, T. vaginalis virus (TVV). However, there are few reports on the properties of the virus. To further determine variations in protein expression of T. vaginalis, we detected 2 strains of T. vaginalis; the virus-infected ($V^+$) and uninfected ($V^-$) isolates to examine differentially expressed proteins upon TVV infection. Using a stable isotope N-terminal labeling strategy (iTRAQ) on soluble fractions to analyze proteomes, we identified 293 proteins, of which 50 were altered in $V^+$ compared with $V^-$ isolates. The results showed that the expression of 29 proteins was increased, and 21 proteins decreased in $V^+$ isolates. These differentially expressed proteins can be classified into 4 categories: ribosomal proteins, metabolic enzymes, heat shock proteins, and putative uncharacterized proteins. Quantitative PCR was used to detect 4 metabolic processes proteins: glycogen phosphorylase, malate dehydrogenase, triosephosphate isomerase, and glucose-6-phosphate isomerase, which were differentially expressed in $V^+$ and $V^-$ isolates. Our findings suggest that mRNA levels of these genes were consistent with protein expression levels. This study was the first which analyzed protein expression variations upon TVV infection. These observations will provide a basis for future studies concerning the possible roles of these proteins in host-parasite interactions.

The First Identified Citrus tristeza virus Isolate of Turkey Contains a Mixture of Mild and Severe Strains

  • Cevik, Bayram;Yardimci, Nejla;Korkmaz, Sava
    • The Plant Pathology Journal
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    • 제29권1호
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    • pp.31-41
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    • 2013
  • The presence of Citrus tristeza virus (CTV) has previously been reported in citrus growing regions of Turkey. All serologically and biologically characterized isolates including I$\breve{g}$d${\i}$r, which was the first identified CTV isolates from Turkey, were considered mild isolates. In this study, molecular characteristics of the I d r isolate were determined by different methods. Analysis of the I$\breve{g}$d${\i}$r isolate by western blot and BD-RT-PCR assays showed the presence of MCA13 epitope, predominantly found in severe isolates, in the I$\breve{g}$d${\i}$r isolate revealing that it contains a severe component. For further characterization, the coat protein (CP) and the RNA-depen-dent RNA polymerase (RdRp) genes representing the 3' and 5' half of CTV genome, respectively, were amplified from dsRNA by RT-PCR. Both genes were cloned separately and two clones for each gene were sequenced. Comparisons of nucleotide and deduced amino acid sequences showed that while two CP gene sequences were identical, two RdRp clones showed only 90% and 91% sequence identity in their nucleotide and amino acid sequences, respectively, suggesting a mixed infection with different strains. Phylogenetic analyses of the CP and RdRp genes of I$\breve{g}$d${\i}$r isolate with previously characterized CTV isolates from different citrus growing regions showed that the CP gene was clustered with NZRB-TH30, a resistance breaking isolate from New Zealand, clearly showing the presence of severe component. Furthermore, two different clones of the RdRp gene were clustered separately with different CTV isolates with a diverse biological activity. While the RdRp-1 was clustered with T30 and T385, two well-characterized mild isolates from Florida and Spain, respectively, the RdRp-2 was most closely related to NZRB-G90 and NZRB-TH30, two well-characterized resistance breaking and stem pitting (SP) isolates from New Zealand confirming the mixed infection. These results clearly demonstrated that the I$\breve{g}$d${\i}$r isolate, which was previously described as biologically a mild isolate, actually contains a mixture of mild and severe strains.

Isolation and Characterization of Chlorella Virus from Fresh Water in Korea and Application in Chlorella Transformation System

  • Park, Hye-Jin;Yoon, Hong-Mook;Jung, Heoy-Kyung;Choi, Tae-Jin
    • The Plant Pathology Journal
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    • 제21권1호
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    • pp.13-20
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    • 2005
  • Chlorella viruses are large icosahedral, plaque-forming, dsDNA viruses that infect certain unicellular, chlorellalike green algae. The genomic DNA of over 300 kb contains many useful genes and promoters. Over 40 chlorella viruses have been isolated from fresh water in Korea since 1998. The viruses were amplified initially in chlorella strain NC64A, and pure isolates were obtained by repeated plaque isolation. SDS-PAGE analysis revealed similar but distinct protein patterns, both among the group of purified viruses and in comparison with the prototype chlorella virus PBCV-1. Digestions of the 330- to 350-kb genomic DNAs with 10 restriction enzymes revealed different restriction fragment patterns among the isolates. The tRNA-coding regions of 8 chlorella viruses were cloned and sequenced. These viruses contain 14-16 tRNA genes within a 1.2- to 2-kb region, except for the SS-1 isolate, which has a 1039-bp spacer in a cluster of 11 tRNA genes. Promoter regions of several early genes were isolated and their activities were analyzed in transformed chlorella. Some promoters showed stronger activity than commonly used CaMV 35S promoter and chlorella transformation vectors for heterologous protein are beings constructed using these promoters.

양식 넙치 치어에서 분리한 birnavirus의 특성 (Characterization of birnavirus isolated from cultured flounder fry)

  • 손상규;박명애;도정완;정초록;박정우
    • 한국어병학회지
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    • 제8권2호
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    • pp.91-98
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    • 1995
  • 최근 남해안 일대의 육상 양식장에서 사육중이던 넙치(Paralichthy olivaceus) 치어가 폐사하여 조사한 결과 3개 양식장에서 바이러스가 분리되었다. 분리된 바이러스들은 모두 외막이 없는 정육면체 모양이었으며 50~55mm 정도의 크기를 지녔다. 전기영동상에서 RNA와 구조 단백질의 patterns를 확인하고, IPNV에 대한 항혈청을 사용하여 중화실험을 수행한 결과, 분리한 세 바이러스는 birnavirus인 IPNV와 매우 유사함이 밝혀졌다. 특히 분리 바이러스중 CS는 IPNV의 AB 혈청형과 DS와 YJ는 SP 혈청형과 유사하였다.

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한국영아에서 분리된 로타바이러스의 VP7 유전자형 및 염기서열 분석 (Typing and Sequence Analysis of the VP7 Gene of Rotavirus Isolated from Infants in Korea)

  • 송미옥;윤여란;정상인;최철순;임인석;강신영;안창남;김원용
    • 대한바이러스학회지
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    • 제30권2호
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    • pp.101-112
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    • 2000
  • Rotaviruses are the most common cause of severe vomiting and diarrhea in children worldwide and classified as a genus in the family Reoviridae. Rotavirus has eleven segmented dsRNAs and the virion consists of three shells. Outer capsid VP7 and VP4 induce neutralizing antibodies and are classified into G types (glycoprotein VP7) and P types (protease-sensitive VP4). Characterization of VP7 gene of Korean isolates of human rotavirus was performed using multiplex PCR and nucleotide sequence analysis. After RT-PCR amplification of full length (1,062 bp) of VP7 genes, the amplified PCR products were G typed by multiplex PCR and the nucleotide sequences were compared with those of reference rotavirus from GenBank. The G type analysis revealed that 25% (2/8) belong to G1, whereas 37.5% (3/8) benong to G2 and G4, respectively. The Korean isolates within the same serotypes showed high homology of nucleotide sequences and could be discriminated from foreign isolates exception with two strains (CAU009 and CAU022). But Korean isolates CAU009 and CAU022 were close related into japanease isolates 417 (99.2%) and indian isolates (97.6%) than Korean isolatese. Our results showed that these two strains were supposed to be originated from abroad. As a results, The G typing and nucleotide sequence analysis of VP7 gene of rotavirus isolated from infants in Korea could be used for identification, serotying and determination of novel or unusual strains of rotaviruses.

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Characterization of Melon necrotic spot virus Isolated from Muskmelon

  • Park, Gug-Seoun;Kim, Jae-Hyun;Kim, Jeong-Soo
    • The Plant Pathology Journal
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    • 제19권2호
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    • pp.123-127
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    • 2003
  • A severe disease of muskmelon (Cucumis melo cv. Alsnight) grown on rockwool in a plastic house was characterized by leaf and stem necrosis followed by death of the plants. In 2001, an isolate of Melon necrotic spot virus-MN (MNSV-MN) of the genus Camovirus was identified as the causal agent of the disease on the basis of biological reactions and nucleotide sequence analyses of coat protein (CP) gene. MNSV-MN induced necrotic local lesions on mechanically inoculated leaves and systemic necrotic spots on the upper leaves of melon cvs. Alsnight, Rui III, Party, Imperial, and Seolhang. However, the inoculated leaves of watermelon and cucumber showed only necrotic lesions. DsRNAs extracted from the melon infected with MNSV-MN were separated into three components. Molecular sizes of the dsRNAs were estimated at approximately 4.5, 1.8, and 1.6 kbp. The amplified cDNA products of CP gene for MNSV-MN by RT-PCR showed approximately 1.2 kbp. The amplified DNA was digested to three fragments by MspI treatment. The cDNA of the genomic RNA of MNSV-MN was cloned and the region deduced to encode the CP was sequenced. The CP coding region, located near 3' end of the genome, consisted of 1,170 nucleotides and had the potential to encode a 390 amino acid protein. The nucleotide and amino acid sequences of MNSV-MN CP gene were 84.0-94.6% and 90.8-94.9% identical with other MNSV isolates found in the GeneBank database, respectively. This is the first report on the occurrence of MNSV in Korea.

Characterization of Melon necrotic spot virus Occurring on Watermelon in Korea

  • Kwak, Hae-Ryun;Kim, Jeong-Soo;Cho, Jeom-Deog;Lee, Joong-Hwan;Kim, Tae-sung;Kim, Mi-Kyeong;Choi, Hong-Soo
    • The Plant Pathology Journal
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    • 제31권4호
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    • pp.379-387
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    • 2015
  • Melon necrotic spot virus (MNSV) was recently identified on watermelon (Citrullus vulgaris) in Korea, displaying as large necrotic spots and vein necrosis on the leaves and stems. The average occurrence of MNSV on watermelon was found to be 30-65% in Hapcheon and Andong City, respectively. Four isolates of the virus (MNSV-HW, MNSV-AW, MNSV-YW, and MNSV-SW) obtained from watermelon plants in different areas were non-pathogenic on ten general indicator plants, including Chenopodium quinoa, while they infected systemically six varieties of Cucurbitaceae. The virus particles purified by 10-40% sucrose density gradient centrifugation had a typical ultraviolet spectrum, with a minimum at 245 nm and a maximum at 260 nm. The morphology of the virus was spherical with a diameter of 28-30 nm. Virus particles were observed scattered throughout the cytoplasm of watermelon cells, but no crystals were detected. An ELISA was conducted using antiserum against MNSV-HW; the optimum concentrations of IgG and conjugated IgG for the assay were $1{\mu}l/ml$ and a 1:8,000-1:10,000 dilutions, respectively. Antiserum against MNSV-HW could capture specifically both MNSV-MN from melon and MNSV-HW from watermelon by IC/RT-PCR, and they were effectively detected with the same specific primer to produce product of 1,172 bp. The dsRNA of MNSV-HW had the same profile (4.5, 1.8, and 1.6 kb) as that of MNSV-MN from melon. The nucleotide sequence of the coat protein of MNSV-HW gave a different phylogenetic tree, having 17.2% difference in nucleotide sequence compared with MNSV isolates from melon.