Cloning and expression of ebola virus GP and VP40 structural proteins
Lee, Carmen Ching Pei
Date of Issue2016-05-31
School of Biological Sciences
Cloning and expression of ebola virus GP and VP40 structural proteins by Lee Ching Pei, Carmen. Ebola virus (EBV) is a member of the Filoviridae family which is made up of seven structural proteins. EBV exists as five different species with varying pathogenicity in human and non-human primates. The recent re-emerging Zaire ebola virus (ZEBOV) outbreak in West Africa caused about 50% of fatality. To date, there is no specific treatment or vaccine available to treat EBV. Recombinant proteins, ZEBOV glycoprotein (GP) and virion protein (VP40), of the virus were used to look into the localisation and effects it had on mammalian cells. To elucidate the localisation and protein expression of GP and VP40, the recombinant proteins were tagged with FLAG or cMyc. GP is the only transmembrane protein found on the virion surface. GP induced cell rounding, detachment and syncytia when transfected with mammalian cells. Based on immunofluorescence assay (IFA) results, protein expression of GP were mainly located at the perinuclear region and plasma membrane of the cells. VP40 is EBV matrix protein and as the name implies, the protein size was found to be 40kDa from western blot result. Data from IFA suggest that VP40 was found in the cells cytoplasm and plasma membrane.
Final Year Project (FYP)
Nanyang Technological University