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AVB101 - E.coli B strain for efficient in vivo biotinylation

Strain AVB101 is an E. coli B strain (hsdR, lon11, sulA1) contains pBirAcm, an engineered pACYC184 plasmid with an IPTG inducible birA gene to over-express the biotin ligase. This glycerol stock of bacteria is shipped frozen.

Quantity: 1 vial, containing 0.5 ml of bacterial glycerol stock.
This product is shipped on dry ice and has to be stored at -80°C.

Strain AVB101, an E. coli B strain (hsdR, lon11, sulA1), contains pBirAcm , an IPTG inducible plasmid containing the BirA gene engineered into pACYC184. It is compatible with most cloning vectors and is maintained with Chloramphenicol (10 μg/ml). This strain is recommended for protein expression because of its robust growth and the absence of the OmpT and Lon proteases.

At the time of induction, Biotin should be added at a concentration of 50 μM. The pAN and pAC vectors containing the AviTagTM require ampicillin (100 μg/ml).

Cells are supplied as a 0.5 ml liquid culture in 30% glycerol. Store at -80°C.

Note: AVB101 cells are not competent. For BirA expressing competent cells please look at related products.

  • What vectors are compatible with AVB101?

    Most expression vectors that are used in laboratories today are derive from the colE1 plasmid which includes pBR322, pUC, pGEM, pBAD and are compatible with AVB101. If you want to avoid a two plasmid system you could also use AVB100 where the BirA encoded on the bacterial chromosome and is expressed via the arabinose promoter.

  • Why is chloramphenicol not added to the growth/induction media to maintain the BIP-300 plasmid when it is used in the initial overnight culture? (relevant only if AVB101 E. coli is used as the host strain)

    We find that the chloramphenicol is not necessary at this point to maintain the pBirAcm BirA over-expression plasmid. It is well enough maintained because of its low copy number.

Ashraf SS, Benson RE, Payne ES, Halbleib CM, Grøn H.

A novel multi-affinity tag system to produce high levels of soluble and biotinylated proteins in Escherichia coli.
Protein Expr Purif. 2004 Feb;33(2):238-45.

Comment: Use of Avitag for in vivo biotinylation of recombinant expressed proteins.

Link to publication