I have a mutant fly line that was generated by inserting a p-element in the 5' UTR of my gene. My RT-PCR suggests the transcript is elevated in my mutant. I am confident in my controls; they look correct. I'm just wondering if anyone has observed this before?
Similar situation has been reported. Please see the paragraph of the attached paper from PNAS: "P Element Insertions Caused Up-Regulation of apontic and pointed genes". I highlighted it in yellow, 2 places.
Hi Malachi,
Yes - We've definitely seen P-element insertions that result in elevated transcript levels...
Hi Malachi,
Although it is possible that the insertion can cause upregulation of gene expression.
But you can find out whether the p-element is an EP line AND whether there is any GAL4 in the background?
Is the RT-PCR done in heterozygous or homozygous background?
Similar situation has been reported. Please see the paragraph of the attached paper from PNAS: "P Element Insertions Caused Up-Regulation of apontic and pointed genes". I highlighted it in yellow, 2 places.
Thank you everyone for your responses. To answer your questions, Sudipta, gal4 is not in the background and this is an EY element. The pcr is performed on embryos from homozygous mutant mothers mated to homozygous mutant fathers. It turns out the increase in transcript is not statistically significant after running three biological replicates. We have decided to take an alternative approach to answer our question. Thanks again!
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