gfp reporter construct

Model Organism: Worms Methods & Reagents
1

Hello,

Sorry for this naive question…

I want to check the expression of my promoter using a gfp reporter. I cloned my promoter upstream of the gfp fragment I PCRed from pCM1.53 (867 bp fragment starting at the ATG, containing 3 introns) to which I added a stop.
Now that I look closer at gfp reporter vectors, I see that the pPD95.75 actually also bears an artificial intron between the MCS and the GFP coding region. Is this intron really necessary? Shall I add it to my construct?
http://www.wormbook.org/chapters/www_reportergenefusions/reportergenefusions.html

Thank you very much for your help.

Audrey

2

It may be worthwhile to add it. I have always included the intron, so I can’t say myself how experiments go leaving it out. If you look a bit further down the page where the authors discuss PCR fusions,

Transcriptional reporters: Fusing a promoter to the MCS of a pPD95.75-type expression vector ensures that the intron between the MCS and reporter gene is included. Inclusion of this intron greatly enhances reporter gene expression.
3

Thanks for your reply!