I've noticed that after transducing my pQCXIP-geneX retrovirus into my cells, I have detectable expression of geneX (by western blot) the following day. Am I just seeing the effects of high expression from only a few cells that had an integration event, or is dsDNA that was reverse transcribed from the viral RNA being transcribed and translated in the absence of integration? A pQCXIP-GFP construct would be useful in answering this question, however I do not have this readily available.
Does anyone have experience with this? I am hoping that retrovirus could be a useful "transient" expression vector that I can use without selection in antibiotics for short-term assays.
Depending on the strength of your antibody, you can pick up extremely minute quantities of protein by WB. That does not necessarily indicate transcription or translation prior to integration. Since the cells you use would be non-synchronised, the integration event could be happening in multiple cells at multiple times.
Using a pQCXIP-GFP may not answer your question either for the same reason here. But, atleast with HIV, there is evidence that Nef is translated from unintegrated DNA. I dont know if it is true of all other proteins or if it is true of any other retrovirus. But, the possibility exists that it can happen.
And as to your last question, you can use a retroviral vector as a transient expression system if you transfect it in. If you use it in a transduction, it gets stably integrated. You may not need to select with antibiotics, depending on your multiplicity of infection.
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