pCBFD1/HBFD1Bad
(Plasmid #53364)

• Purpose: Will convert beta-carotene into astaxanthin in E. coli when expression of the hbfd1 cDNA is induced with arabinose. Use with pAC-BETA or pAC-BETAipi.
• Depositing Lab: Francis X Cunningham Jr
• Publication: Cunningham FX et al Plant Cell. 2011 Aug;23(8):3055-69. doi: 10.1105/tpc.111.086827. Epub 2011 Aug 23.

Backbone

• Vector backbone: pHBFD1Bad
• Backbone manufacturer: Francis X. Cunningham, Jr.
• Backbone size w/o insert (bp): 5271
• Total vector size (bp): 6707
• Modifications to backbone: To make pCBFD1/HBFD1Bad, the cbfd1 cDNA was excised from pCBFD1 as a PvuII-XhoI fragment and ligated into pHBFD1Bad that had been digested with KpnI, blunted with the Klenow enzyme, purified by phenol/chloroform extraction, and then digested with XhoI and gel purified.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Growth instructions: For production of astaxanthin, introduce this plasmid into E. coli strain TOP10 that already contains plasmid pAC-BETA or pAC-BETAipi, induce expression of the hbfd1 cDNA with arabinose (0.1% is satisfactory), and grow in liquid culture on a platform shaker at 28 degrees Celsius in darkness for 2-3 days.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: cbfd1
• Alt name: AdKeto1
• Alt name: carotenoid β-ring 4-dehydrogenase
• Species: Adonis aestivalis
• Insert Size (bp): 1457
• GenBank ID: AY644757.1
• Promoter: lac

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: PvuII (destroyed during cloning)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: none
• 3′ sequencing primer: none

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry