• Journal of Microbiology and Biotechnology
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• v.27 no.12
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• pp.2173-2179
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• 2017

The intracytoplasmic membrane of Rhodobacter sphaeroides readily vesiculates when cells are lysed. The resulting chromatophore membrane vesicle (CMV) contains the photosynthetic machineries to synthesize ATP by ATPase. The light-dependent ATPase activity of CMV was lowered in the presence of $O_2$, but the activity increased to the level observed under anaerobic condition when the reaction mixture was supplemented with ascorbic acid (${\geq}0.5mM$). Cell lysis in the presence of biotinyl cap phospholipid (bcp) resulted in the incorporation of bcp into the membrane to form biotinylated CMV (bCMV), which binds to streptavidin resin at a ratio of approximately $24{\mu}g$ bacteriochlorophyll a/ml resin. The ATPase activity of CMV was not affected by biotinylation, but approximately 30% of the activity was lost by immobilization to resin. Interestingly, the remaining 70% of ATPase activity stayed constant during 7-day storage at $4^{\circ}C$. On the contrary, the ATPase activity of bCMV without immobilization gradually decreased to approximately 40% of the initial level in the same comparison. Thus, the ATPase activity of CMV is sustainable after immobilization, and the immobilized bCMV can be used repeatedly as an ATP generator.

• The Plant Pathology Journal
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• v.23 no.4
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• pp.251-259
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• 2007

Two isolates of Cucumber mosaic virus (CMV) originated from lily plants, named Ly2-CMV and Ly8-CMV, were compared with their pathological features in several host plants. Ly2-CMV and Ly8-CMV could induce systemic mosaic symptom in Nicotiana benthamiana, but Ly2-CMV could not systemically infect tomato and cucumber plants that have been used for CMV-propagative hosts. While Fny-CMV used as a control infected systemically the same host plants, producing typical CMV symptoms. Ly8-CMV could infect systemically two species of tobacco (N. tabacum cv. Xanthi-nc and N. glutinosa) and zucchini squash (Curcubita pepo), but Ly2 failed systemic infection on these plants. As resulted from tissue-print immunoblot assay, different kinetics of systemic movement between Ly2-CMV and Ly8-CMV were crucial for systemic infection in tobacco (cv. Xanthi-nc). Sequence analysis of full-length genome of two lily isolates showed Ly2 and Ly8 belonged to subgroup IA of CMV. The lily isolates shared overall 98 % sequence identity in their genomes. Coat protein, 3a protein, and 2b protein involved in virus movement was highly conserved in genomes of the isolates Ly2 and Ly8. Although there is the low frequency of recombinants and reassortants in natural CMV population, phylogenetic analysis of each viral protein among a number of CMV isolates suggested that genetic variation in a defined population of CMV lily isolates was stochastically produced.

• Korean Journal Plant Pathology
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• v.2 no.3
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• pp.145-149
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• 1986

A tomato Aspermy Virus (TAV-B) served as a helper virus for multiplication and encapsidation of satellite RNAs which were isolated from two different CMV isolates, D and K. These two satellite RNAs induced renarkable attenuation of TAV symptoms in infected tobacco, which was correlated with a reduction of virus content in the plant. The CMV satellite RNAs also caused lethal necrosis in TAV-infected tomato as in the case of CMV system.

• Korean Journal of Radiology
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• v.21 no.6
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• pp.726-735
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• 2020

Objective: Recent innovations in biology are boosting gene and cell therapy, but monitoring the response to these treatments is difficult. The purpose of this study was to find an MRI-reporter gene that can be used to monitor gene or cell therapy and that can be delivered without a viral vector, as viral vector delivery methods can result in long-term complications. Materials and Methods: CMV promoter-human organic anion transporting polypeptide 1B3 (CMV-hOATP1B3) cDNA or CMV-blank DNA (control) was transfected into HEK293 cells using Lipofectamine. OATP1B3 expression was confirmed by western blotting and confocal microscopy. In vitro cell phantoms were made using transfected HEK293 cells cultured in various concentrations of gadoxetic acid for 24 hours, and images of the phantoms were made with a 9.4T micro-MRI. In vivo xenograft tumors were made by implanting HEK293 cells transfected with CMV-hOATP1B3 (n = 4) or CMV-blank (n = 4) in 8-week-old male nude mice, and MRI was performed before and after intravenous injection of gadoxetic acid (1.2 µL/g). Results: Western blot and confocal microscopy after immunofluorescence staining revealed that only CMV-hOATP1B3-transfected HEK293 cells produced abundant OATP1B3, which localized at the cell membrane. OATP1B3 expression levels remained high through the 25th subculture cycle, but decreased substantially by the 50th subculture cycle. MRI of cell phantoms showed that only the CMV-hOATP1B3-transfected cells produced a significant contrast enhancement effect. In vivo MRI of xenograft tumors revealed that only CMV-hOATP1B3-transfected HEK293 tumors demonstrated a T1 contrast effect, which lasted for at least 5 hours. Conclusion: The human endogenous OATP1B3 gene can be non-virally delivered into cells to induce transient OATP1B3 expression, leading to gadoxetic acid-mediated enhancement on MRI. These results indicate that hOATP1B3 can serve as an MRI-reporter gene while minimizing the risk of long-term complications.

• Korean Journal of Organic Agriculture
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• v.19 no.spc
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• pp.271-274
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• 2011

Pepper mild mosaic virus (PMMoV) and cucumber mosaic virus (CMV) are important pathogens in various vegetable crops worldwide. We have found that methanol extracts of Quercus dentate (Oaimyo Oak) gal/nut strongly inhibit PMMoV and CMV infection. Based on this result, the inhibitor named as "KN0912" formulated from the extract of Q. dentate gallnut was tested for its inhibitory effects on PMMoV or CMV infection to each local lesion host plant (Nicotiana glutinosa; PMMoV, Chenopodium amaranticolor; CMV). Pre-treatment effect of KN0912 against infections of each virus to local host plant was measured to be $75.1{\pm}0.5{\sim}97.5{\pm}1.5%$ to PMMoV and $70.6{\pm}2.2{\sim}99.0{\pm}1.0%$ to CMV in 1~10mg/ml conc. and the absorption effect of the antiviral composition of KN0912 to the inside of tobacco leaves tissue, was inhibited by 55.7% to PMMoV and 63.8% to CMV. The persistence of KN0912 treatment was maintained until after the 3 days high inhibitory effect by 98% to PMMoV and by 95.1% to CMV. Inhibitory effects on systemic host plants of KN0912 were measured to be 80~90% to PMMoV and 60~75% to CMV. From the change of morphological characteristics of PMMoV particles under EM, we are tentatively suggested that one mode of action of KN0912 is inactivation due to the destruction of virus particles.

• Research in Plant Disease
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• v.17 no.2
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• pp.211-215
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• 2011

Cucumber mosaic virus (CMV)-like isolate was collected from Raphanus sativus (cv. Choon-hyang), which showed mosaic symptoms. The isolate was confirmed to a strain of CMV by host responses in Vigna unguiculata, Chenopodium amaranticolor and Gomphrena globosa, by viral genome composition with RT-PCR and PCR-RFLP, and by serological analysis. Symptom developed by the strain of CMV was severe in Nicotiana benthamiana, N. glutinosa, N. tabacum (cv. Samsun, cv. Xanthi), Cucumis melo (cv. Early hanover), Cucumis sativus (cv. White wonder), Capsicum annuum (cv. Chung-yang and cv. Geum-top), but mild symptom was developed in Raphanus sativus (cv. Choon-hyang), Brassica rapa ssp. pekinensis (cv. Bul-Am No. 3), and B. juncea (cv. Daenong Jukgot). Newly isolated strain of CMV could infect diverse crops including Solanaceae, Cucurbitaceae and Brassicaceae. We designated the new strain of CMV as Gn-CMV based on the novel infectivity of Brassicaceae. In double-stranded (ds) RNA analysis, Gn-CMV consisted of 3.3, 3.0, and 2.2 kb genomes likewise other strains of CMV. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) showed 28 kDa of the CMV coat protein. By restriction enzyme mapping using Cac8I, ClaI and MspI of RT-PCR products indicated that Gn-CMV belongs to CMV subgroup I.

• The Plant Pathology Journal
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• v.28 no.4
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• pp.381-389
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• 2012

Molecular characterization of Cucumber mosaic virus (CMV) was done by using samples from tomato and cucurbitaceous plants collected from different locations in the northwest region of Iran. After screening by enzyme-linked immunosorbent assay, 91 CMV-infected samples were identified. Biological properties of eight representative isolates were compared with each other revealing two distinct phenotypes on squash and tomato plants. Phylogenetic analyses based on nucleotide sequences of the coat protein (CP), movement protein (MP) and 2b of the new isolates, together with that of previously reported isolates, led to the placement of the Iranian isolates in subgroups IA and IB according to CP and MP genes, but in subgroup IA according to the 2b gene. These data suggest that reassortment may have been a major event in the evolution of CMV in Iran, and that the Iranian isolates are derived from a common recent ancestor that had passed through a bottleneck event.

• BMB Reports
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• v.30 no.4
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• pp.269-273
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• 1997

Heptocvte-specific expression induced by Hepatitis B virus (HBV) enhancer 2-core gene promoter was examined in various hepatocyte and non-hepatocyte cell lines. using non-viral and retroviral vector systems in which chloramphenicol acetyltransferase (CAT) is used as a reporter. The non-viral plasmid containing the HBV enhancer 2-core promoter exhibited 22 and 66% of CAT activities in hepatoma cell lines. HepG2 and Hep3B, respectively when compared with CAT activity expressed by CMV promoter. The CAT activities, however. were found to be marginal in other tested hepatoma cell lines as well as mouse primary hepatocytes and non-hepatocytes. The HBV enhancer 2 located upstream the CMV promoter did not affect the CMV promoter activity nor provided hepatocyte-specific expression. Transfection of retroviral plasmid DNA containing the HBV enhancer 2-core promoter as an internal promoter exhibited high and specific CAT expression in HepG2 and Hep3B cell lines but the activity value was 5 to 10 fold lower than the non-viral plasmid with identical promoter. These results suggest that the usage of HBV enhancer 2-core promoter for liver specific expression is limited to certain vectors and hepatocyte cell lines.

• Korean journal of applied entomology
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• v.20 no.2 s.47
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• pp.76-82
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• 1981

Cucumber Mosaic Virus (CMV) was isolated from naturally infected Hippenstrum hybridum. The virus caused mosaic symptoms on Nicotiana glutinosa and local lesions on Vignaunguiculata. The thermal inactivation point was 56C, dilution end point $10^{-3}$ and longevity in vitro was 2 days for CMV from Hippeastrum. Purified virus was obtained using citrate chloroform extraction procedure and polyethylene glycol precipitation followed by sucrose density gradient centrifugation. Purified virus had a typical absorption at 245nm. Electron micrographs of the purified virus from Hippeastrum showed spherical particles with 30nm in diameter. The purified virus reacted with CMV antiserum in agar gel double diffusion test.

• Toxicological Research
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• v.20 no.2
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• pp.109-116
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• 2004

For the safety evaluation of adenovirus-mediated gene transfer, we investigated differential gene expressions after transfecting adenoviral vector containing p16 tumor suppressor gene (Ad5CMV-p16) into human non-small cell lung cancer cells. In the previous study, we showed adenovirus-mediated $p16^{INK4a}$ gene transfer resulted in significant inhibition of cancer cell growth. We investigated gene expression changes after transfecting Ad5CMV-p16, Ad5CMV (null type, a mock vector) into A549 cells by using cDNA chip and oligonucleotide microarray chip (1200 genes) which carries genes related with signal transduction pathways, cell cycle regulations, oncogenes and tumor suppressor genes. We found that $p16^{INK4a}$ gene transfer down regulated 5 genes (cdc2, cyclin D3, cyclin B, cyclin E, cdk2) among 26 genes involved in cell cycle regulations. Compared with serum-free medium treated cells, Ad5CMV-p16 changed 27 gene expressions, two fold or more on oligonucleotide chip. In addition, Ad5CMV-p16 did not seem to increase the tumorigenicity-related gene expression in A549 cells. Further studies will be needed to investigate the effect of Ad5CMV-p16 on normal human cells and tissues for safety evaluation.