Mutations were engineered in E, NS3 and NS4B protein genes to improve replication in Vero cells.
The mutations were engineered both at the donor (L131L, M160L, M197F and M202H) and acceptor (M265I and M234E) sides.
HCM-associated JPH2 mutations were engineered and functionally characterized using immunocytochemistry, cell morphometry measurements, and live cell confocal calcium imaging.
Mutations were engineered by Quick-change mutagenesis (Agilent Technologies) in a pcDNA3.1-Kv7.2 pcDNA3.1-Kv7.2lectroplasmidogical and western-blot experiments), or in a dual-tagged Enhanced Green fororelectrophysiological-hemandlutinin (EGFP-Kv7.2-HA) plasmid (for immunocytochemistry experiments), as described21,34,61.
To assess their effect on antiviral activity of Oas1b, the NS3 and 2K mutations were engineered into an infectious WNV cDNA clone.
Mutations were engineered into the VLP genome to produce particles deficient in activities associated with viral reverse transcriptase, RNase H, and RNA packaging.
As part of a general study into the impact of quaternary structure on enzyme function, a library of 31 point mutations were engineered at the dimer dimer interface of the homotetrameric 8-barrel protein, N-acetylneuraminate lyase (NAL, EC 4.1.3.3).
To test the consequences of subtle structural alterations remote from the RdRp active-site, the following single point mutations were engineered in the WNV NS5 RdRp coding region: T363N, A365N, and T537I; these mutations were selected in an effort to stabilize the secondary structural elements near the rNTP binding pocket of the RdRp.
These mutations are engineered into a recombinant genome that is inserted into a cell, where it generates a recombinant virus.
Point mutations were engineered using the QuikChange XL site-directed mutagenesis kit (Stratagene).
