Hello,

There are different versions of GFP or Fluorescent proteins with sequence modifications.

How do know Valine has changed ?? How did you sequence the protein ? or just predicted ?

Its true that Valine is 2nd A.A.

Just design siRNA and it should work.

I have not came accross PTMs on GFP.

https://www.uniprot.org/uniprot/C5MKY7

https://www.addgene.org/13031/sequences/

Good luck

Anand

Hello Anand Patwardhan,

Thank you very much for your reply. I just "guessed" from the Addgene software sequence that the 2nd aminoacid (valine) was not present in the final version of EGFP since it is marked in a different way in the sequence, as I show in the picture attached.

Valine is shown as the aminoacid with number "1a" instead of "2" as it should be since it's the second aminoacid and it looks like it is not included in the EGFP aminoacid sequence.

What does the tag "1a" on valine mean then? Thank you again for your time.

Kind regards,

Adrian

Hello Adrian Dario Juncos Bombin

Dont worry, there is no Post translation modification here..

Most Methionine are removed, hence Valine could become new 1st Amino acid.

Check these discussions

https://www.researchgate.net/post/Does_every_protein_start_with_methionine

Just design siRNA and you should be fine.

Regards

Anand