pAC-ZEAX
(Plasmid #53274)

• Purpose: Contains crtE, crtB, crtI, crtY and crtZ genes of Erwinia herbicola (Pantoea agglomerans) Eho10, and thereby produces zeaxanthin in E. coli
• Depositing Lab: Francis X Cunningham Jr
• Publication: Sun et al J Biol Chem. 1996 Oct 4;271(40):24349-52.

Backbone

• Vector backbone: pAC-EHER
• Backbone manufacturer: Francis X. Cunningham, Jr.
• Backbone size w/o insert (bp): 13581
• Total vector size (bp): 12478
• Modifications to backbone: The plasmid pAC-ZEAX was constructed from pAC-EHER by deletion of a 1.1-kb SalI-SalI fragment containing the Erwinia herbicola Eho10 gene (crtX) encoding zeaxanthin glucosyltransferase. The C terminal end of the idi gene was also deleted, although this was not done purposefully. The function of the product of the idi gene (ORF6), and the salutary effect of IDI on carotenoid production in E. coli were not recognized until later.
• Vector type: low copy number bacterial cloning vector

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Top10
• Growth instructions: Grow liquid cultures on a platform shaker at 28 degrees Celsius for 2-3 days in darkness for best zeaxanthin production, or grow on agar plates at room temperature for 3-7 days.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: crtE, crtY, crtI, crtB, crtZ
• Species: Erwinia herbicola Eho10
• Mutation: Deleted idi (ORF6) and crtX genes
• GenBank ID: M87280.1
• Promoter: endogenous promoters

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NA (unknown if destroyed)
• 3′ cloning site: NA (unknown if destroyed)
• 5′ sequencing primer: none
• 3′ sequencing primer: none

Resource Information

• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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

Depositor Comments

For better yield of low copy number pAC-based plasmids, grow liquid cultures on a platform shaker at ca. 30 degrees Celsius. When cultures reach early stationary phase, dilute 2-fold with growth medium, add spectinomycin (150 mg/liter), and "amplify" for several hours before harvest. For plasmid selection and maintenance in E. coli, use chloramphenicol at 30 mg/liter.