My objective is to purify protein from insect cells but I suspect a problem with the bacmid. Upon transfection of the cells with non-recombinant bacmid using cellfectin, cells do undergo morphological changes such as detachment, increase in size of cell and diameter and formation of occlusion bodies but there is no cell lysis. Even on keeping the plate for 7-10 days, only 10% of 40-50% transfected cell lyse. The presence of baculoviruses was checked by TEM but there were none.I had thoroughly checked bacmid preparation on gel and PCR before transfection.
I'm not quite sure what you are asking, but typically in our lab we do not see much change in the cell morphology etc after transfection. It is only after further amplifying the virus that we observe lysed cells. So we would take a sample of the supernatant from the transfection plate, use this to infect a new culture in shake flask and then look for any changes (after 5-7 days). Do you have another bacmid you can use as a positive control? This would help you troubleshoot your procedures.
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