I have been using Tk- cells for vaccinia virus infection. They were working fine till all of a sudden, shortly after adding the virus they lift off. I tried everything including changing the PBS and the plates i am using but no luck any idea why ?
Hi Sherief,
Don’t worry, it’s a typical feature. Information was obtained with thymidine kinase-deficient LM cells, and virus-induced thymidine kinase has also been studied in system infected with vaccinia virus. It was found that enzyme synthesis ceases at a certain stage of the viral cycle, while virus-induced RNA synthesis in IHD-infected LM(TK-) cells was not completely suppressed, and cells treated with actinomycin D before infection were still able to produce about 40% of the normal amount of thymidine kinase, as assayed 7 h after infection or earlier. Also, others investigated this point in more detail and found that in contrast to other systems described earlier, the arrest of IHD-induced thymidine kinase synthesis does not require viral DNA replication. Data presented suggested that the expression of an early genetic function could be responsible for arrest of the enzyme synthesis.
Best wishes,
Ilya
Hi Ilya
Thank you for your reply but this is not really the problem I am having.
The problem is that without even adding any virus to the cells, just adding PBS will result in the cells to lift off. They never did that before
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