Expressing an SSB protein

Dr. Artem Evdokimov eudokima at mail.ncifcrf.gov
Mon May 7 17:24:13 EST 2001


Did you mean purification of specifically SSB's or any old protein in
general ?

As far as expression systems go, we love recombinational cloning from
LifeTech but that's us :)

A.

martin murray wrote:
> 
> We have identified a gene in an unculturable bacterium which appears
> to code for a single-stranded DNA binding protein. It is part of a
> larger plasmid fragment and has been cloned into pUC18. We are
> interested in possibly expressing this gene, purifying the protein,
> and determining whether it will bind SS DNA in vitro.
> Not having done this before, or anything like it, we have some
> questions:
> 
> Which of the many available expression kits would be best for this
> experiment?
> 
> Is it necessary to cut out the relevant ~400 bp ORF and reclone that
> into the expression vector,
> or can we use the pUC18 construct we already have?
> 
> What is involved in the protein purification process?
> 
> What conditions are required for the actual DNA-binding step?
> 
> I have looked for answers to these questions on the internet but
> haven't had much luck, and there
> is no one at this facility to ask. Thanks very much for any advice you
> can give.
> 
> MM

-- 
|Dr. Artem Evdokimov   Protein Engineering |
| NCI-Frederick        Tel. (301)846-5401  |
|              FAX (301)846-7148           |
|        eudokima at mail.ncifcrf.gov         |
|      http://www.ncifcrf.gov/plague       |




More information about the Methods mailing list