Write 4 pages thesis on the topic foot and mouth malady virus. From this paper, it is clear that the primary long-term goal of the project is to determine the interactions of release factors specifically needed for the 2A reaction. The method for determining this is genetical to isolate mutations within genes responsible for encoding release factors in the yeast cell that pause the 2A response.
Upon synthesis of ubiquitin as part of a polyprotein, it is removed from polyprotein by specific cytosolic proteinases, indicating a new N-terminus to the remainder of the protein. Arginine shows clearly that this protein is degraded urgently as soon as it is degraded. It follows that the stability of an Ade2p portion of the protein depends on the activeness of 2A peptide. The white color of the yeast generates a color-based selection for mutants.
i.) PCR was used to amplify SUP45 (encoding the yeast eRF1) from a plasmid template under conditions that involves the reduction in the fidelity of the polymerase hence generating random point mutations within the gene.
ii.)Integrated reporters that only allow assessment of both stop codon read-through and 2A actions into a sup45Δ, pCEN-URA3-SUP45 yeast strain.
iii.) Transformation of the PCR product and a linearized low plasmid copy possessing sequences homologous at the terminals. The underlying of PCR product into underlying yeast strain from existing (2) thus developing through the in-vivo recombination