Efficient Biotinylation E. coli B Strain AVB101

Product Description

Strain AVB101 is an E. coli B strain (hsdR, lon11, su1A1), containing a pACYC184 plasmid with an IPTG-inducible birA gene to overexpress biotin ligase (pBirAcm). This strain is recommended for protein expression because of its robust growth and the absence of the OmpT and Lon proteases.

Expression of both biotin ligase and the AviTag protein are induced with IPTG (1 mM). Biotin should be added at the time of induction to a concentration of 50 µM. The pAN and pAC vectors require ampicillin for maintenance and the recommended concentration is 100 µg/mL.

Features of plasmid pBIRAcm

• Backbone is pACYC184.
• 1.3 Kb fragment contains lacIq inserted at EcoRV site (thus disrupting tetR gene).
• BirA is expressed from the tac promoter operator.
• rrnB terminator construct is between the laqIq region and destroys XbaI site of pACYC184.