Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Animal and Molecular Model Systems > Arabidopsis and Plant Biology
Register Search Today's Posts Mark Forums Read

Arabidopsis and Plant Biology Discuss Arabidopsis and Plant Biology Research.


A question about studying plant gene subcellular localization byusing eYFP

A question about studying plant gene subcellular localization byusing eYFP - Arabidopsis and Plant Biology

A question about studying plant gene subcellular localization byusing eYFP - Discuss Arabidopsis and Plant Biology Research.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 01-29-2007, 04:21 PM
Chan Kam Ho
Guest
 
Posts: n/a
Default A question about studying plant gene subcellular localization byusing eYFP



Dear all,

I have tried to study the subcellular localization of my gene by fusing it
in frame with eYFP and also DsRed2. I have placed it under the control of
CaMV and I have sequenced the constructs and I am sure that the construct is
correct and there is no misake in the construct. I tried to overexpress the
construct in transgenic Arabidopsis and since my construct also contains
CaMV GUS I found that my transgenics are GUS positive but I can't see the
fluorescence of eYFP/ DsRed2. Is it possible that the fluorescent proteins
doesn't fold correctly when it is fused with my protein of interest and so I
can see the fluorescence? does anyone have similar experience to share? What
can I do now?


Thanks in advance
Kam Ho
Reply With Quote
  #2  
Old 01-29-2007, 11:01 PM
Oliver Berkowitz
Guest
 
Posts: n/a
Default A question about studying plant gene subcellular localizationbyusing eYFP

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

--
_______________________________

Dr. Oliver Berkowitz
The Australian National University
Research School of Biological Sciences
Environmental Biology Group
GPO Box 475
Canberra ACT 0200
Australia

P: +61 - (0)2 - 61254549
F: +61 - (0)2 - 61254919

[Only registered users see links. ]

"Chan Kam Ho" <[Only registered users see links. ]> wrote in message
news:[Only registered users see links. ].net...
is
the
I
What


Reply With Quote
  #3  
Old 01-30-2007, 12:32 PM
John Runions
Guest
 
Posts: n/a
Default A question about studying plant gene subcellular localizationby using eYFP


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][Only registered users see links. ].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:[Only registered users see links. ].uk
2. [Only registered users see links. ]
3. [Only registered users see links. ]
Reply With Quote
Reply

Tags
byusing , eyfp , gene , localization , plant , question , studying , subcellular


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Human Cytome Project - Update 24 Jan. 2005 Peter Van Osta Cell Biology and Cell Culture 1 08-01-2010 02:18 PM
New Saccharomyces Sequences 11/27/04 Mike Cherry Yeast Forum 0 11-29-2004 12:39 AM
New Saccharomyces Sequences 09/08/04 SGD Sequences Yeast Forum 0 09-13-2004 10:07 PM
New Saccharomyces Sequences 08/11/04 SGD Sequences Yeast Forum 0 08-12-2004 12:26 AM
New Saccharomyces Sequences 05/19/04 SGD Sequences Yeast Forum 0 05-23-2004 04:06 PM


All times are GMT. The time now is 11:22 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.17152 seconds with 16 queries