Abstract
Background: Epidemics and pandemics have had a great impact on the course of human history. As such emergence and re-emergence of diseases of high risk to the individual, and/or the community, are of specific interest and concern to public health systems, whether they be in developed or developing countries. Ebola viruses represent prime examples of such emerging pathogens. Ebola outbreaks are unpredictable, with high severity and fatality rates. This high case fatality rate of Ebola virus infection and the lack of approved vaccines and therapeutics resulted in classification of Ebola viruses as biosafety level 4 pathogens and potential bioweapons by the WHO and the CDC. The main goal of this work is to identify epitopes within the viral proteins of Sudan ebolavirus (strain Gulu) that are targets of the humoral immune response in survivors. Epitope specific antibodies are important indicators of survival from this deadly disease and can serve as templates for the development of synthetic or vectored vaccines
Methods: Using standard cloning techniques the Sudan ebolavirus (strain Gulu) genes for all seven structural proteins were cloned and sequenced. For viral protein 24 (VP24), 30 (VP30), 40 (VP40) and NP further expression constructs were generated containing a Flag tag epitope for detection. These proteins were expressed in 293T cell using standard techniques and transfection efficiency was controlled by using a VP40-GFP fusion protein construct.
Following expression, viral proteins were isolated and tested by western blot analysis.
Methods: Using standard cloning techniques the Sudan ebolavirus (strain Gulu) genes for all seven structural proteins were cloned and sequenced. For viral protein 24 (VP24), 30 (VP30), 40 (VP40) and NP further expression constructs were generated containing a Flag tag epitope for detection. These proteins were expressed in 293T cell using standard techniques and transfection efficiency was controlled by using a VP40-GFP fusion protein construct.
Following expression, viral proteins were isolated and tested by western blot analysis.
Original language | English |
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Pages (from-to) | e461-e462 |
Journal | International Journal of Infectious Diseases |
Volume | 14 |
Issue number | SUPPLEMENT 1 |
DOIs | |
State | Published - 8 Mar 2010 |