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

Abstract

OBJECTIVE:

In order to compare the infection of HIV-1 pseudovirus to suspended and adherent cells, Tzmbl cells containing beta-gal (beta-galactosidase) reporter gene were used here to do the analysis. 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, beta-gal positive Tzmbl cells and virus infection were determined using X-gal staining and beta-glo (beta-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 single-round replicative pseudovirus in the future.

• Authors: Yan Su, Lingling Su, Tao Yin, Baojiang Zhang
• Journal: Wei sheng wu xue bao = Acta microbiologica Sinica (Wei Sheng Wu Xue Bao) Vol. 52 Issue 7 Pg. 921-6 (Jul 04 2012) ISSN: 0001-6209 [Print] China
• PMID: 23115978 (Publication Type: Comparative Study, English Abstract, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Suspensions
• beta-Galactosidase

Topics

• Cell Adhesion
• Genes, Reporter (genetics)
• HEK293 Cells
• HIV-1 (physiology)
• Humans
• Suspensions
• Transfection
• Viral Load (genetics)
• Virus Replication (genetics)
• beta-Galactosidase (genetics)