Hi all,
I am transfecting HEK293T cells with SARS-Cov-2 spike protein. I will likely be transfecting the entire sequence after I clone my insert into my plasmid. However, most publications do not clone the first twelve aa residues and add a C9/C5 signal sequence to the plasmid so that the insert will follow it. Do I have to add this non-native signal sequence, or should I not omit the first 12 residues and just utilize the native signal sequence. I am aiming for high scale protein production. Thanks!
You can insert the constructed sequence from the plasmid to the desired cell through heat shock or other mechanisms.
Brett Seese What is the reason provided in literature for removing those sequences? The signal sequence can be upstream with an omission of a few residues of ORF for fusion, but what is the importance of removing 12 residues?
This study reported the expression cassette: You can follow the same:
Article Native-like SARS-CoV-2 spike Glycoprotein expressed by ChAdO...
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