I'm looking to tag endogenously some proteins in Leishmania major, and since I'm trying to repeat something done before in T. brucei using the YFP tag, I was planning to do the same in L. major. Has anyone used or are aware of work done in Leishmania using endogenously tagged proteins with YFP? I am looking for constructs used to do this, and was not successful finding literature on it, as it is all in T. brucei. Is there any specific reason on why it has not been used in Leishmania? Or, does anyone think there would be a difference in using GFP rather than YFP (there's work done in Leish using GFP)? I assume they have a very similar structure?
Dear Catarina,
I hope these papers may help you to answer your questions. In my experience I had no problem when using G or YFP, but I did it in episomal over expressing plasmids, which is not exactly your case. Even knowing that both GFP and YFP share similar structures, some details such as fluorescence stability are not exactly the same, being YFP more interesting.
http://www.pnas.org/content/106/18/7583.full
http://www.jbc.org/content/285/36/27982.full
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267249/
http://ec.asm.org/content/11/5/662.full
Best wishes!
Oi Rubens,
Thanks for the reply, I'll take a look then! I'm more tempted to try YFP since I'll try to do in Leish what was done in Tryps.
Obrigado!
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