Difference between revisions of "PGP1331"

From SubtiWiki
Jump to: navigation, search
m (moved Pgp1331 to PGP1331)
Line 3: Line 3:
 
'''General comments'''
 
'''General comments'''
  
Integrative Plasmid for the fusion of 3x FLAG tag to the C-terminus of a protein, keeping the control of the expression under its natural promotor
+
Integrative plasmid for the fusion of 3x FLAG tag to the C-terminus of a protein, keeping the control of the expression under its natural promotor
*E.coli: multicopy plasmid, select for ampicillin
+
* ''E.coli'': multicopy plasmid, select for ampicillin
*B. subtilis: no ori, select for spectinomycin
+
* ''B. subtilis'': no ori, select for spectinomycin
  
 
'''Hints for the usage'''
 
'''Hints for the usage'''
*amplify the last 600bp of your gene of interest,  
+
* amplify the last 600bp of your gene of interest,  
*skip the Stopp-Codon of the gene in the revers primer
+
* skip the Stopp-Codon of the gene in the revers primer
*take care that the integration is in-frame
+
* take care that the integration is in-frame
  
 
'''Antibody'''
 
'''Antibody'''
 
*dilute the anitbody 1:10000 (you can freeze the Blotto solution and reuse it)
 
*dilute the anitbody 1:10000 (you can freeze the Blotto solution and reuse it)

Revision as of 13:22, 16 March 2010

pBQ200: click to enlarge

General comments

Integrative plasmid for the fusion of 3x FLAG tag to the C-terminus of a protein, keeping the control of the expression under its natural promotor

  • E.coli: multicopy plasmid, select for ampicillin
  • B. subtilis: no ori, select for spectinomycin

Hints for the usage

  • amplify the last 600bp of your gene of interest,
  • skip the Stopp-Codon of the gene in the revers primer
  • take care that the integration is in-frame

Antibody

  • dilute the anitbody 1:10000 (you can freeze the Blotto solution and reuse it)