I have been trying to get a sample of bacteriophage CE6 (which expresses T7 polyemerase) for protein expression studies. The lysogenic form of this is the (DE3) you see in genotypes of cells used for expression from T7 promoters. I have a need for the 'phage itself.
Can anyone send me a small sample as the commercial supplier has spent over six months trying to deal with my order.
EMD-Millipore (formerly Novagen) sells a titer phage lysate. I don't think that CE6 is exactly the same phage as used to create the DE3 lysogen. I believe CE6 is designed to not integrate at all so that it can be used for infection of plasmid containing cells, whereas the DE3 is designed to be a stable lysogen. You can probably recover some DE3 phages directly from any existing DE3 lysogen (like BL21 (DE3)) but be sure to use an R- permissive host.
thanks Michael.
I had both the phage and the lysogenisation kit once, but they have since disappeared. I used the lysogenisations kit before, never the phage. Now I want to control two promoters (T7 and Tac) differentially and I thought the CE6 infection trick would be one way of doing it.
Unfortunately its Novagen who are having trouble making it. Over seven months now from my initial order and they are still having trouble.
Have you considered changing one of your promoters, perhaps recloning one of the inserts so that instead of the Tac promoter you are using a different inducer, arabinose comes to mind for example although there are many others.
it was easier (I thought) just to purchase the 'phage !!!
They want nearly €400 for it... which I have probably lost from my research fund as its the end of the fiscal year. Groan..
This was supposed to be a quick 'see if it changes anything' idea s I wanted to co-express two proteins. I have done it various ways before but we have the plasmid clones already etc etc.
thanks for the suggestion though.
- Badges
- Science method
- Similar topics
- Biological Science
- Genetics
- Gene Expression
- Protein Expression