| Bacterial Strains |
| Strain AVB 100 |
| AVB100 is an E. coli K12 strain [MC1061 araD139 delta(ara-leu)7696 delta(lac)l74 galU galK hsdR2(rK-mK+) mcrB1 rpsL(Strr)] with 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, pAN, pATN and pATC AviTag vectors allows independent control over the expressed gene of interest and the BirA levels. |
| Use of this strain is limited to site of purchase. Please review the license included before using this product. |
| Strain AVB99 |
Strain AVB101 |
| Strain AVB99 is an E.coli strain (XL1-Blue) containing a pACYC184 plasmid with an IPTG-inducible birA gene to overexpress biotin ligase (pBirAcm). The plasmid should be maintained with chloramphenicol at 10 µg/mL concentration (this is a low copy-number plasmid). |
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. |
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| 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. |
| Plasmid in Avidity's AVB99 & AVB101 |
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- The backbone is pACYC184.
- 1.3 Kb fragment contains lacIq inserted at EcoRV site (thus disrupting tetR gene).
- The BirA is expressed from the tac promoter operator.
- The rrnB terminator construct is between the laqIq region and destroys XbaI site of pACYC184.
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