Plasmodium falciparum: recombinant baculoviruses direct the expression of circumsporozoite proteins in Spodoptera frugiperda cell cultures
| Autor: | Alex Bollen, Omer van Opstal, Pierre Voet, Paul Jacobs, Fabienne Milican, Dirck Gheysen, Pascal Gilles, Michel Heinderyckx, Marc Massaer |
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| Rok vydání: | 1991 |
| Předmět: |
Baculoviridae
Recombinant Fusion Proteins Genetic Vectors Molecular Sequence Data Plasmodium falciparum Protozoan Proteins Antibodies Protozoan Sf9 Antigens Protozoan Spodoptera Moths law.invention Mice law parasitic diseases Genetics Animals Molecular Biology Cells Cultured biology Base Sequence Isoelectric focusing Immunodominant Epitopes fungi General Medicine DNA biology.organism_classification Molecular biology Circumsporozoite protein Cell culture Recombinant DNA Plasmids |
| Zdroj: | Molecular biology reports. 15(2) |
| ISSN: | 0301-4851 |
| Popis: | The DNA coding for the circumsporozoite protein (CPS) of Plasmodium falciparum has been cloned into the baculovirus expression vector pAcYM1 and expressed in Spodoptera frugiperda (Sf9) insect cells. Three DNA constructs have been made: the first one directs the synthesis of the complete CSP (aa 1-412), the second leads to the production of a species devoid of the anchor domain (aa 1-391) and the third one to a molecule lacking both signal and membrane anchor sequences (aa 18-391). All three recombinant CPS were produced at about 3 micrograms per 10(6) infected cells and were characterized in terms of immunoreactivity and apparent molecular weight. Analytical purification of the recombinant proteins was achieved by a combination of heat treatment, acidification, isoelectric focusing and ion exchange chromatography. The purified material, when injected into mice, generated only modest antibody responses, although antisera from immunized mice reacted with control CSP antigens carrying or not the major immunodominant repeat region. |
| Databáze: | OpenAIRE |
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