pSV7d-Env plasmid病毒Env表达质粒-BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

WT-Env and SOS-Env expression.

pSV7d expression vectors bearing SOS-Env or wild-type Env (WT-Env) genes (JR-FL strain). Env was expressed in HEK 293T cells by cotransfecting with the HIV-1 rev-bearing pCDNA3 plasmid (Invitrogen, Carlsbad, Calif.) at a 2:1 Env/Rev ratio, with a standard calcium phosphate method. Surface expression of SOS-Env and WT-Env was quantified by flow cytometry with monoclonal antibody 2G12 and fluorescein isothiocyanate-conjugated goat anti-human immunoglobulin secondary antibody. The fusion proteins were expressed at low densities through the pSV7d plasmid. SOS-Env was almost completely but not totally proteolytically cleaved into 120-kDa and 40-kDa subunits, as observed in reducing gels. In nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the uncleaved and the cleaved but disulfide-linked SOS-Env cannot be distinguished because they migrate with the same mobility.

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