Fusion recombinant protein expression of newcastle disease virus from Escherichia coli-Cloned C1a using accurapidTM protein expression kit

Abstract

This research aimed to express F recombinant protein that is clon from genes F of local isolate ND virus which can be used as vaccine candidate in order to improve the effectiveness of ND virus vaccination. Confirmation of NDV pBT7-N-His-Fusion plasmid on C1a clone is done by gel agarose 1 % electrophoresis with staining by using fluorosafe DNA stain. To separate plasmid and insert that contain genes F, cutting is done with EcoRI restriction enzyme. EcoRI enzyme is able to cut NDV pBT7-N-His-Fusion plasmid through 37°C incubation process during three hours. DNA cutting visualization is done by gel agarose 1% electrophoresis by using fluorosafe DNA stain. NDV pBT7-N-His-Fusion plasmid is express by E. coli extract in order to gain F protein. The product of protein expression is visualized by SDS-PAGE and western blot. NDV pBT7-N-His-Fusion plasmid visualization by gel agarose electrophoresis results 4643 bp band. Moreover, from the visualization of Eco RI enzyme cutting on gel agarose electrophoresis result, the researcher found two bands with different size, 4001 bp and 642 bp. After protein expression process 25,6 kDa band is seen both in the result of SDS-PAGE and western blot.