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Versatile Reporter Vectors for Monitoring Viral Transduction

3.0 x 107

2.8 x 106

pFB-Neo-LacZ
vector

1:102

5.0 x 106

1:103

6.0 x 106

* Titer = (fraction of cells fluorescing over background) x (supernatant dilution/ml) x (2 x 105 cells infected). Titers for pFB-Neo-LacZ vector = (fraction blue cells / total # cells in field) x (supernatant dilution / ml) x (2 x 105 cells infected); titers represent the average for 3 randomly chosen fields for each dilution.

Fig.2

pFB-Neo-LacZ Vector

The use of b-galactosidase as a reporter in retroviral vectors has a number of advantages over other reporters. The enzymatic activity of b-galactosidase can be used as a quantitative measure of gene expression from the viral promoter in a cell line of choice. In addition, virally transduced cells can be visualized in situ by staining with X-gal, and viral titers may be determined both by counting blue cells using light microscopy or by FACS analysis of infected cells following incubation with a fluorescent b-galactosidase substrate. Additionally, by employing the pFB-Neo-LacZ vector, viral titers may also be determined by G418-resistant colony formation due to the presence of the neo m
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