Dear Kam and others,
Oliver's suggestion to use transient expression techniques when
evaluating fluorescent protein expression from your vectors is a good
one. As well as particle bombardment techniques, we use a
transient-expression-in-tobacco-leaves trick that is much simpler and
quicker than stably transforming arabidopsis. This has just been
published as Sparkes et al (2006) Nature Protocols 1: 2019-2025. If
you would like a pdf, let me know.
Cheers, John.
Oliver Berkowitz wrote:
Hi Kam,
had similar problems with my constructs (in pAVA vectors) changing to
another vector with a different linker between GFP and my cDNA worked. Also,
you might try transient expression by particle bombardment as you can
increase the amount of DNA you shoot and thus the expression of your
construct.
Cheers
Oliver
--
*********************************
C. John Runions, Ph.D.
School of Life Sciences
Oxford Brookes University
Oxford, UK
OX3 0BP
email: [1]jrunions At brookes.ac.uk
phone: +44 (0) 1865 483 964
web: [2]http://www.brookes.ac.uk/lifesci/runions/HTMLpages/index.html
New - Oxford Brookes Master's in [3]Bioimaging with Molecular
Technology
References
1. mailto:jrunions At brookes.ac.uk
2. http://www.brookes.ac.uk/lifesci/runions/HTMLpages/index.html
3. http://www.brookes.ac.uk/go/bioimaging_masters
