Retroviral vector and GFP

Alex Chang lpss at unixg.ubc.ca
Fri Nov 20 19:38:30 EST 1998


Dear netters,

I have a LNCX based retroviral vector which was used for expressing 
GFP. BOSC cells were used for packaging and NIH 3T3 cells used for 
viral titration.

The transient transfection by Superfect of BOSC was great (90%
efficiency and highy expression level). However, after 48-72 hrs
infection of 3T3 cells by the packaged retroviral vector, the GFP
expression level was very low (you can barely see them under
fluorescent microscope). Eventhough, about 80-90% cells were
infected. 

The retroviral infection was done using the 48 hr harvested viral 
supernatant, which was centrifuged and polybrene mixed (4 ug/ml).

Any clue?

Alex Chang
Pathology
University of British Columbia
alex at cmmt.ubc.ca



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