Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Molecular Biology Techniques
Register Search Today's Posts Mark Forums Read

Molecular Biology Techniques Molecular Biology Forum. Includes forums for common molecular biology techniques.


help needed with prokaryotic expression

help needed with prokaryotic expression - Molecular Biology Techniques

help needed with prokaryotic expression - Molecular Biology Forum. Includes forums for common molecular biology techniques.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 02-26-2009, 01:32 AM
Volunteer
Points: 1,873, Level: 26 Points: 1,873, Level: 26 Points: 1,873, Level: 26
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2009
Posts: 33
Thanks: 0
Thanked 4 Times in 3 Posts
Default help needed with prokaryotic expression



Hi,

I tried to express a segment about 600 bp inserted in pET30 in BL21 strain. The ORF is correct by sequencing, however, the protein is not expressed at 30 or 37 degree with 0.1 to 1 mM IPTG induction.

I appreciate very much for any suggestions.
Reply With Quote
  #2  
Old 02-26-2009, 12:21 PM
Pipette Filler
Points: 3,020, Level: 35 Points: 3,020, Level: 35 Points: 3,020, Level: 35
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Apr 2006
Location: Bavaria, Germany
Posts: 18
Thanks: 0
Thanked 4 Times in 3 Posts
Arrow Re: help needed with prokaryotic expression

How do you determine whether or not your PoI is expressed?

I didnot find the pET30 vector; I do find the pET-30a/b/c on Addgene. What I see, is the ORF regulated by Lac operon is in the anti-clockwise direction. Are you sure that your insert is not reverted?

best and good luck
Reply With Quote
Reply

Tags
expression , needed , prokaryotic


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
Human Cytome Project - an idea - Update 19 April 2005 Peter Van Osta Cell Biology and Cell Culture 1 06-01-2009 02:17 PM
qPCR NEWSLETTER - October 2006 Editor www.Gene-Quantification.info Protocols and Methods Forum 0 10-26-2006 12:25 PM
A Human Cytome Project - an idea - Update 14 March 2005 Peter Van Osta Cell Biology and Cell Culture 0 03-14-2005 02:27 PM
Human Cytome Project - Update 6 Jan. 2005 Peter Van Osta Cell Biology and Cell Culture 0 01-06-2005 11:18 AM


All times are GMT. The time now is 05:43 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.16682 seconds with 16 queries