I would like to ask about the transfection of A549 cells. I used pCMV6-entry plasmid to overexpress one gene of interest (I want to study its role in inhibiting influenza virus replication) in alveolar epithelial cells (A549 cells). I used empty vector as a control and Lipofectamine as mock treatment, but I found that even the empty vector could reduce the influenza virus replication as much as my complete construct do. My question is, does anybody work with pCMV6-entry plasmid and have some induction of cytokines that could also attenuate the virus replication? As I want to know the phenotype that I got due to the overexpression of my gene of interest or due to the plasmid artifact. Thanks in advance.
Thanks Andrei Nikonov for your replay, Actually I am using qiagen miniprep kit. Checking the plasmid at different conc. is a good idea and I am trying now to use different vector.
Best regards
Mohamed
No I do not think so. but I know that my gene of interest is highly up-regulated by Endotoxins, so If I got Endo contamination, I would see this in the empty vector (something that I did not get so far), meaning that I do not have such contamination of Endotoxins from my mimiprep kit.
Thanks a lot Randy, I included also mock treated (lipofectamine) in my experiment and it seems that it has not anyeffect. Thanks again for the paper
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