Method: The SARS-CoV-2 Wuhan-1
spike, cloned into pCDNA3.1, was mutated using the QuikChange Lightning Site-Directed Mutagenesis kit (Agilent Technologies) and NEBuilder HiFi DNA Assembly Master Mix (NEB) to include
D614G (wild-type) or lineage defining mutations for Beta (
L18F,
Result: Additional substitutions include
P25L (in ~43% of viruses) and
W152R (in ~7%) in the NTD,
T478K (~17%) in the RBM,
L585F (~17%) in S1,
P681H (~8%) adjacent to the furin cleavage site,
A879T (~7%),
D936H (~5%), and
H1101Q (~8%) in S2, with additional amino acid changes detected in less than 5% of viruses.