The reaction product was then treated using the QuikChange II XL Site-Directed Mutagenesis Kit (Stratagene, CA, US), according to the manufacturer's instructions.
Phagemid pCADS was constructed by mutagenizing the TAG amber stop codon of pCAD phagemid by CAG codon with the primer SupASC using the QuikChange II Site-Directed Mutagenesis Kit (Agilent Technologies) following the manual instructions.
The Ser47Ala, Thr530Ala and stop mutation sites were mutated by using the QuikChange II XL Site-Directed Mutagenesis Kit.
TRIM5αrh point mutants were generated using the QuikChange II XL site-directed mutagenesis kit (Stratagene, Cedar Creek, TX).
Site-directed mutagenesis was performed using the Quikchange II site-directed mutagenesis kit (Stratagene) according to manufacturer's instructions.
All point mutations were generated using the QuikChange II XL site-directed mutagenesis kit according to the manufacturer's recommendations (Agilent Technologies).
The mutant constructs pF/H426-UBP2-C745S and pF/H426-RUP1-Y135F, bearing site specific point mutations were constructed using the QuikChange II Site-Directed Mutagenesis kit (Stratagene).
Plasmids containing single point mutations and the RSLESD deletion construct were prepared using the QuikChange II XL Site-Directed Mutagenesis Kit (Stratagene, Amsterdam, The Netherlands).
