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Expression of porcine circovirus 2 ORF2 gene requires codon optimized E. coli cells

The result's identifiers

  • Result code in IS VaVaI

    <a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62157124%3A16170%2F07%3A00001431" target="_blank" >RIV/62157124:16170/07:00001431 - isvavai.cz</a>

  • Result on the web

  • DOI - Digital Object Identifier

Alternative languages

  • Result language

    angličtina

  • Original language name

    Expression of porcine circovirus 2 ORF2 gene requires codon optimized E. coli cells

  • Original language description

    Expression and purification of whole and NLS deleted ORF2 capsid protein of porcine circovirus 2 is demonstrated in the present study. Gene coding for both protein forms were cloned into pDest17 vector and expressed in BL21 (DE3)AI cells and in BL21-CodonPlus (DE3)-RIPL E. coli cells. The later cells were used to overcome difficulties with the heterologous expression of viral proteins in prokaryotic systems. Whole 30 kDa recombinant ORF2 protein was successfully expressed in BL21-CodonPlus (DE3)-RIPL cells only, 3 mg of pure protein was consistently obtained per liter of bacterial culture. NLS deleted ORF2 protein was expressed in both cell types. Resulting proteins reacted with PCV2 positive swine serum in immunofluorescent test and immunoblot.

  • Czech name

    Exprese ORF2 porcinního cirkoviru2 vyžaduje E. coli buňky s optimalizací kodonů

  • Czech description

    V práci byla popsána exprese a purifikace ORF2 kapsidového proteinu porcinního cirkoviru 2. Gen kódující protein byl klonován do pDest17 vektoru a exprimovány v BL21 (DE3)AI a in BL21-CodonPlus (DE3)-RIPL E. coli buňkách. 30 kDa rekombinantní ORF2 protein byl úspěšně exprimován v BL21-CodonPlus (DE3)-RIPL buňkách a byly získány , 3 mg čistého proteinu na litr bakteriální kultury.

Classification

  • Type

    J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

  • CEP classification

    EE - Microbiology, virology

  • OECD FORD branch

Result continuities

  • Project

    <a href="/en/project/GA524%2F06%2F1640" target="_blank" >GA524/06/1640: The control of viremia by neutralizing antibodies and localization of neutralization epitopes of porcine circovirus 2</a><br>

  • Continuities

    P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Others

  • Publication year

    2007

  • Confidentiality

    S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Data specific for result type

  • Name of the periodical

    Virus Genes

  • ISSN

    0920-8569

  • e-ISSN

  • Volume of the periodical

    34

  • Issue of the periodical within the volume

    2

  • Country of publishing house

    US - UNITED STATES

  • Number of pages

    6

  • Pages from-to

    199-204

  • UT code for WoS article

  • EID of the result in the Scopus database