mutant GFP - green phenotype of E coli

[Mega [Andreas Stuermer]]

Hi guys! 

I was wondering if anyone knows where to get the sequence of the mutant GFP that makes E coli look green even under daylight... 

http://diyhpl.us/~bryan/papers2/diybio/Carolina_transformations_manual.pdf (page 10) 

[Mega [Andreas Stuermer]]

Does someone even have it or sequenced it?

[Koeng]

I think it's called sfGFP. That's the most effective one. 

Or, you could do what I used to do, which is just keep it constantly expressed and grow the cells for much longer. Worked for me as well

I also have that plasmid that they use if you would like a sample. Ironically, it has the GFP gene regulated by the lac operator yet still expresses well under normal conditions. Probably because there are more plasmid copies than lac repressor proteins in the cell, but I've never really investigated it.

-Koeng

[Mega [Andreas Stuermer]]

Thanks! You are right it seems.
http://parts.igem.org/Part:BBa_I746916

Its in-vivo properties are considerably improved with respect to mut3 - it develops fluorescence about 3fold faster than mut3 GFP and reaches 4fold higher absolute fluorescence levels. Fluorescenct colonies can be identified with the naked eye even without UV or blue light illumination (that is to say the amount of blue light in normal daylight or lablight is sufficient). Additionally it is more stable in vitro and refolds faster after in vitro denaturation with respect to mut3 GFP.

[Josiah Zayner]

Most GFPs I have worked with make bacteria look green under normal lighting conditions. It more depends on the quantity of the GFP than having a specific GFP. Same is true for RFP and the likes.

[Koeng]

In my experience, RFP is more visible than GFP under normal lighting conditions. May want to use that instead

-Koeng

[Andreas Stuermer]

Ok, great to know, thanks!!

--
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diy...@googlegroups.com. To unsubscribe from this group, send email to diybio+un...@googlegroups.com. For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to a topic in the Google Groups "DIYbio" group.
To unsubscribe from this topic, visit https://groups.google.com/d/topic/diybio/P2i39ueT950/unsubscribe.
To unsubscribe from this group and all its topics, send an email to diybio+un...@googlegroups.com.
To post to this group, send email to diy...@googlegroups.com.
Visit this group at http://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/e10d8a7d-8d79-4fc6-b6a4-a44f2b6fd399%40googlegroups.com.
For more options, visit https://groups.google.com/d/optout.