i was wondering if i have to include a unc-54 3’UTR region (or similar) for proper expression of a reporter protein other than GFP.
i’m curretly planning the construction of a expression vector with DENDRA2 as the reporter protein…
any help is appreciated.
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Whether you use GFP, or any other fluorescent protein you need 3’UTR. Ideally the endogenous 3’UTR region should be used, but Unc54 will do.
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BE careful when using unc-54 3-ÚTR.
In our hands, we detected unespecific GFP expression in the most-posterior intestinal cells which was prevented when using the gene-specific 3´-UTR.
This unespecific GFP expression in the intestine depends very much on the lenght of the promoter or promoter+gene insert you use. The longer usually the lower unspecific GFP signal
Hope it helps
Antonio Miranda Vizuete, PhD.
Centro Andaluz de Biología del Desarrollo (CABD-CSIC)
Dpto. de Fisiología, Anatomía y Biología Celular
Universidad Pablo de Olavide
Carretera de Utrera Km. 1
Teléfono +34 954 348558
Fax +34 954 349376