HRS and VPS4 cDNA were cloned into pEGFP-C vector (Clontech, France). The VPS4 E223Q mutant was made by PCR mutagenesis using the QuikChange II site directed mutagenesis kit (Stratagene, France). Expression vectors for HRS fused to the Streptavidin binding protein (SBP) and Calmodulin binding protein (CBP) affinity tags were obtained by cloning HRS into the pNTAP-B vector (Stratagene, France). The expression vector for Flag-tagged BST-2 was obtained by cloning of BST-2 into p3XFlag vector. cDNA for HA tagged-Vpu cloned into pCDNA3.1 was a gift from Dr. Florence Margottin-Goguet [45] All mutagenesis and subclonings were verified by DNA sequencing.
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