Hello Fellows,
I want to clone a protein with a Flag but I can't decide where should I place the Tag on C-terminal or N-terminal since some papers put it on N-terminal but others put on C-terminal ?
The protein is a transcription factor ATF3.
Anyone can give me a clue.
Thank you :)
Hi
Dr Doaa
pl find the answer here
Producing a Flag-Tagged Protein - Biologicalworld.com
biologicalworld.com/flagtag.htm
Protocol on how to combine a flag tag with a protein to produce a flag-tagged protein. ... If attaching the Flag tag to the C-terminus of the protein, remember to remove ... If attaching the Flag tag to the N-terminus, it is probably better to attach it ...
ajit
Hello, Doaa.
If the target is a transmembrane or GPI-linked or organellar protein to be delivered to specific organelles, care should be taken to consider the position of signal peptide within the TF. Have a look where is signal peptide in your TF, N- or C-termini. If it hasn't got any NLS at both termini, it's OK. I would not worry about the TF too much even if it hasn't got NLS because other guiding proteins will do the nuclear translocation job. However, it would be a better idea to have both clones of plasmids during cloning for rainy day.
Regards.
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