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.