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Expression of Biologically Active Measles Virus Hemagglutinin Glycoprotein by a Recombinant Baculovirus

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Author(s): Z.A. Sadigh | M.S. Shahrabadi | A. Shafyi | B. Bambai

Journal: Pakistan Journal of Biological Sciences
ISSN 1028-8880

Volume: 11;
Issue: 9;
Start page: 1220;
Date: 2008;
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Keywords: Measles virus AIK-C strain | hemagglutinin | recombinant protein | pDONR221

ABSTRACT
In this study, one of the measles virus membrane proteins, named hemagglutinin (H) which has a key role in tropism, receptor binding, hemagglutinating activity and also induction of protective immunity against viral infection, was expressed by the baculovirus expression system using specific plasmid (pDONR221) to produce entry clone. Measles Virus (AIK-C strain) genome was extracted from infected Vero cells. H gene was amplified by specific primers during RT-PCR reaction and inserted into the specific plasmid (pDONR221) using BP recombination reaction. Recombinant baculovirus harboring H gene was consequently constructed by LR reaction. Insect cells (Sf9) were infected with recombinant baculovirus. In order to increase viral titer, recombinant baculoviruses were passaged four times in Sf9 cells. Synthesis of H protein was verified by SDS-PAGE, western-blot and indirect immunoflourescene using goat polyclonal antibody against Measles Virus. The results showed that H protein was partially glycosylated, but it appeared to be active in hemagglutination assay.
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