Expression of Dengue Viral Proteins in Drosophila Cells
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AbstractDengue virus causes dengue fever, a debilitating illness marked by high fever, headache, muscleand joint pain, and rash. The more severe forms of the illness, dengue hemorrhagic fever and dengueshock syndrome, are life threatening and are a principal cause of hospitalization among children inSoutheast Asia. Globally, the four serotypes of dengue are responsible for up to 100 million infectionsannually, and with the introduction of the WP albopictus vector mosquito to North America, dengue nowposes a threat to the mainland United States. We will develop a safe and effective subunit vaccineagainst dengue infection. The Phase I research will test the feasibility of expressing dengue structuralproteins in a Drosophila cell culture system and examine their association in a lipoprotein particle.Co-expression of the two membrane- anchored structural proteins of a related flavivirus have beenshown to yield highly immunogenic proteins with native conformations in a particle; the Drosophila cellexpression system has expressed high levels (30 mg/L) of recombinant HIV gpl2O with biologicalproperties indistinguishable from viral gpl20. High level Drosophila cell expression of dengue proteinsin a lipoprotein particle form could lead to development of a successful tetravalent dengue subunitvaccine.
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