Reading Reagents
Many recombinant baculoviruses and
E.coli strains are constructed as part of our research work. We make particular
use of in-house vectors that are not available commercially to tag
proteins for purification and secretion. Some recombinants we make express proteins of
more general interest and we supply these as purified preparations
to third parties on a cost recovery basis. For example we have
provided HIV reagents to the AIDS reagent repository at NIBSC (CFAR)
over a number of years. Reading reagents is the section of the lab
that deals with this part of our work. If you make use of this
service you are, in essence, buying the know-how of a group of
people who are very experienced at what they do.
Services include:
High throughput recombinant baculovirus
formation. We use the modified baculovirus DNA described by us
(Zhao et al) to ensure rapid recovery of 100% recombinant virus. You
either provide a transfer vector or have us construct one for you
based on the vector suite approach we have developed that is described in
the projects pages. We return to you a guaranteed volume of known
titre virus or use the virus to go forward for large scale infection.
Large scale infections. We run the Wave
BioReactor (www.wavebiotech.com)
at 5L scale for large scale infections as part of a collaboration
with the Oxford Protein Production Facility on the development of
streamlined sources of recombinant protein for structural analysis.
We also use small scale high density cultures based
around 24 well deep blocks and the Glascol Digital
Pulse Incubator/Shaker.
Protein purification.
We do NOT
run traditional soft gel chromatography but we do use a variety of affinity
tags fused to the protein of interest. Current tags we favour are
His, Fc and TAP. We also routinely make GFP fusions which can help
to resolve expression problems and sometimes offers an additional novel tag
for analysis.
Virus archiving. A simple service that
grows, titrates and stores your precious recombinants in a variety
of long term storage conditions. When you need them, viruses (or
E.coli strains) are recovered, tested and dispatched to you. We
will carry out QC tests if you need them (e.g. we will PCR
check the recombinant locus to confirm there has been no virus
degeneration (deletions do occur in the recombinant virus genome
if good virology practices have not been followed see
Pijlman et al).
To use these facilities
contact Sylvia Rua.
Users to date include:
