Using reporter gene to compare infection efficiency of HIV-1 pseudovirus to suspended and adherent cells

Abstract:

Abstract:［Objective］In order to compare the infection of HIV-1 pseudovirus to suspended and adherent cells，Tzmbl cells containing β-gal ( β-galactosidase ) reporter gene were used here to do the analysis． ［Methods］ HIV-1 pseudoviruses were generated by co-transfection of 293T cells with the plasmid pNL43R-E- and HIV envelope expressing plasmid． Supernatant of co-transfected 293T cells was collected and used to infect Tzmbl cells with or without trypsin treatment． Forty-eight hours after infection，β-gal positive Tzmbl cells and virus infection were determined using X-gal staining and β-glo (β-galactosidase) assay． ［Results］The efficiency of HIV pseudoviruses infection of suspended Tzmbl cell was higher than that of adherent cells and the increase of infection correlated with the pseudoviral subtype．［Conclusion］This study may provide a useful method for HIV biological study and neutralization assays using a singleround replicative pseudovirus in the future．