unc-54 3-prime UTR necessity in expressing reporter proteins other than GFP

Model Organism: Worms Methods & Reagents
1

Hi,

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.

thx

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2

Hey,
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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3

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
41013 Sevilla
ESPAÑA (SPAIN)

Teléfono +34 954 348558
Fax +34 954 349376
email amirviz@upo.es