In vivo Biotinylation-Strains

STRAIN AVB101
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. Avidity, LLC provides the strain in three forms- glycerol stock (AVB101), chemically competent cells (CVB101), and electrocompetent cells (EVB101).

Protocol: Induction of bacterial strains for in vivo biotinylation of AviTag™ proteins

Download the specification sheets:
Glycerol stock AVB101
Electrocompetent cells EVB101
Chemically competent cells CVB101.pdf

In vivo Biotinylation with Bacterial Strains
Sequence for pBirAcm
pBirAcm.txt
NOTE The BirA gene is on the complementary strand between 2829-3791 bp
AVB101 diagram (pdf file for printing)

BIO-500 Biotin solution 10 mL 5 mM d-biotin solution (10 mM Bicine, pH 8.3)
STRAIN AVB99
Dnase deficient K-12 E. coli for isolating pBirAcm plasmid
Strain AVB99 is a K12 E. coli strain (genotype: recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1) containing pBirAcm, an engineered pACYC184 plasmid with an IPTG inducible birA gene to over-express the biotin ligase. The endA1 nuclease deficient strain is recommended for isolation of the pBirAcm plasmid that can be used to transform other E. coli strains to a biotin ligase over-expressing strains. The pBirAcm plasmid should be maintained and selected with chloramphenicol at 10 µg/ml concentration (remember this is a low copy number plasmid). Avidity, LLC provides glycerol stocks (AVB99)

Download the specification sheet:
Glycerol stock AVB99

STRAIN AVB100-integrated birA gene strain
AVB100, an E. coli K12 strain [MC1061 araD139 delta (ara-leu)7696 delta(lac)l74 galU galK hsdR2(rK-mK+) mcrB1 rpsL(StrR)], is a birA gene stably integrated into the chromosome. Overexpression of the BirA protein is accomplished by induction with L-arabinose. The stably integrated birA gene does not require antibiotics to be maintained, and use of AVB100 with IPTG-inducible vectors such as Avidity's pAC or pAN. AviTag vectors allows independent control over the expressed gene of interest and the BirA levels. Avidity, LLC provide glycerol stocks (AVB100) or electrocompetent cells (EVB100).

Please note: Strain AVB100 or derivatives when purchased conveys a limited use license to the end user. This license prohibits the purchaser from selling, assigning or transferring this product to any third party without the express written consent of Avidity, L.L.C. Please review this license on the specification sheet below before purchasing this product.

Download the specification sheets:
Glycerol stock AVB100
Electrocompetent cells EVB100
Related products BIP-300 positive control plasmid BIP-300
BIP-300 is a plasmid containing N-terminal AviTagged ß-galactosidase (ß-gal) gene. Plasmid can be used as a positive control for expression in AVB101 or AVB100