Difference between revisions of "PGP1331"

From SubtiWiki
Jump to: navigation, search
Line 1: Line 1:
 
 
'''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'''

Revision as of 14:16, 16 March 2010

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)