CFP vector in fire kit

Model Organism: Worms Methods & Reagents
1

Hi All,

Does anyone know if there is a CFP vector backbone available in the fire vector kit? I am planning to tag my gene of interest with CFP marker.

Thanks
Ritika

2

Hi

Yes. Plasmid 1665: L4663 from Fire kit at Addgene (also known as pPD133.48) it contains worm introns for better expression and nuclear localization signal. But this old CFP is no the best thing in terms of brightness, have that in mind

Alejandro

3

Hi Alejandro,

Thanks for your response. Is there any other CFP vector backbone which you think I could use for this purpose that has an improved brightness?

Ritika

4

Currently there are two better versions of CFP: Cerulean and mTFP1 (Teal)

According to the literature , the % of relative brightness compared to EGFP of these proteins is :

CFP 39
Cerulean 79
mTFP1 162

http://www.microscopyu.com/articles/livecellimaging/fpintro.html

I haven’t found any Cerulean or mTFP1 with worm introns available (maybe someone else in the forum knows or has it?) I’m in the process of generating a gateway-compatible worm mTFP version myself. But again, if your gene is expressed in relatively high levels, maybe just Cerulean or mTFP1 will do the job.

Alejandro

5

We made a “wormized” monomeric Cerulean in John White’s Lab and would be happy to share if needed.