Production of polyclonal antibodies to a recombinant coat protein of potato virus Y

Folwarczna, Jitka; Plchová, Helena; Moravec, Tomáš; Hoffmeisterová, Hana; Dědič, P.; Čeřovská, Noemi

doi:10.1007/s12223-008-0067-1

Cited by 15 publications

(14 citation statements)

References 15 publications

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“…Alfalfa mosaic virus (AMV) polyclonal antiserum was used for the detection of virus in soybean plants in USA [28]. Our results with CP are in agreement with the results obtained for antibodies raised against recombinant Watermelon bud necrosis virus [29], Potato mop-top virus [30], Potato virus Y [31], Potato virus X [31].…”

Section: Discussionsupporting

confidence: 84%

Expression and Production of Polyclonal Antibodies against Recombinant Coat Protein of Peanut bud necrosis virus

Sivaprasad¹,

Bv²,

Sujitha³

et al. 2015

J Plant Pathol Microbiol

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Section: Discussionsupporting

confidence: 84%

Expression and Production of Polyclonal Antibodies against Recombinant Coat Protein of Peanut bud necrosis virus

Sivaprasad¹,

Bv²,

Sujitha³

et al. 2015

J Plant Pathol Microbiol

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“…The cDNA containing either CP or TGBp1 was obtained by immunocapture-reverse transcriptionpolymerase chain reaction (IC-RT-PCR) using anti-PVM IgG, and specific primers for fragments containing CP or TGBp1 (Table 1). The products were cloned into the pTZ57R⁄T vector (Fermentas) and sequenced (Folwarczna et al 2008). Primers were designed for the cloning of desired sequences into pET-45b(+) (Novagen, Darmstadt, Germany) containing 6x His-tag sequence ( Table 1).…”

Section: Methodsmentioning

confidence: 99%

Production of Polyclonal Antibodies to the Recombinant Potato virus M (PVM) Non‐structural Triple Gene Block Protein 1 and Coat Protein

Čeřovská

Moravec

Plchová

et al. 2012

Journal of Phytopathology

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The genes encoding the coat protein (CP) and triple gene block protein 1 (TGBp1) of Potato virus M (PVM) were cloned into expression vector pET‐45b(+) (N‐terminal 6xHis tag) and expressed in E. coli Rosetta gami‐2(DE3). The purified recombinant antigens were used for raising polyclonal antibodies. The antibodies against recombinant CP were successfully used in Western blot analysis, plate‐trapped ELISA and DAS‐ELISA as a coating for PVM detection in infected potato leaf samples. The antibodies against recombinant non‐structural protein detected the TGBp1 only in Western blot analysis. This is the first report of the production of polyclonal antibodies against recombinant coat protein and TGBp1 of PVM and their use for detecting the virus.

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“…The extraction protocol that we used to extract soluble protein from bacteria was from Folwarczna et al (21) and was optimized for our usage. For Transmission electron microscope (TEM) studies, the bacterial cells that were harvested by centrifugation, resuspended in 1 mL of TE, and incubated at 20°C for 5-10 minutes with lysozyme (1 mg/mL).…”

Section: Transmission Electron Microscopymentioning

confidence: 99%

Expression of Cucumber Mosaic Virus Coat Protein and Its Assembly into Virus Like Particles

Rostami¹,

Bashir²,

Pirniakan³

et al. 2014

Biotech Health Sci

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Background: Cucumber mosaic virus (CMV) has isometric particles with a diameter of about 28-29 nm. A useful method to recognize regions from mutagenesis in the coat protein (CP) gene are important for particle assembly and the expression of viral in CP expression systems like bacteria such as E.coli or yeast as well as their assembly into virus-like particles (VLP). Objectives: In this paper, we report the expression and assembly of VLP in cells of E. coli expressing the CMVCP gene. Materials and Methods: In this study, the CMV-CP gene was released from a previously prepared cloning vector. Then, the CMV-CP was ligated into the expression vector. Sequencing was done by Marcrogen, Inc. (South Korea). A recombinant plasmid was transferred to E.coli isolate Rosetta. After inducing by isopropyl thiogalactosides, the molecular weight of the expressed protein was determined by SDS-PAGE. The extraction of proteins was done by NATURE method to see the possible presence of CMV-like particles. Results: CMV-CP was detected by Western blotting by a CMV specific polyclonal antibody and conjugate. The protein extracted from the CP producing clone was studied under a JEOL 100-CXII transmission electron microscope with 100000× magnification at an acceleration voltage of 100 kV. Conclusions: The results showed that the CP gene was expressed in the prokaryotic system successfully and was assembled into the CMVlike particle.

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Production of polyclonal antibodies to a recombinant coat protein of potato virus Y

Cited by 15 publications

References 15 publications

Expression and Production of Polyclonal Antibodies against Recombinant Coat Protein of Peanut bud necrosis virus

Expression and Production of Polyclonal Antibodies against Recombinant Coat Protein of Peanut bud necrosis virus

Production of Polyclonal Antibodies to the Recombinant Potato virus M (PVM) Non‐structural Triple Gene Block Protein 1 and Coat Protein

Expression of Cucumber Mosaic Virus Coat Protein and Its Assembly into Virus Like Particles

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