I am attempting to express a protein using an Adenovirus vector. I have successfully inserted the gene of interest into the Adenovirus and used linear pAdeno to transfect the 293A cell line. However, I did not observe any obvious cytopathic effect (CPE) in the 293A cells after the transfection. To investigate further, I decided to pass the product blindly for another two passages, washing the cells three times with PBS one hour after infection. In the supernatant of the second passage, I was able to detect my gene of interest using PCR. This raises the question of whether the pAdeno carried over to the second passage or if the Adenovirus carrying my gene of interest was able to propagate in the 293A cells. I am puzzled as to why I did not observe any CPE or detachment of cells from the culture plate if the Adenovirus was able to survive.
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