pAC-LYC plasmid產番茄紅素質粒菌種 BioVector NTCC質粒載體菌種細胞基因保藏中心

pAC-LYC plasmid Contains 3 genes (crtE, crtI, and crtB) of the carotenoid pathway gene cluster of Erwinia herbicola (Pantoea agglomerans) Eho10 and thereby produces lycopene in Escherichia coli
? Vector backbone pAC-EHER
? Backbone size w/o insert (bp)13581
? Total vector size (bp) 9476
? Modifications to backbone
Constructed from pAC-EHER by deletion of adjacent 0.8 and 1.1 kB BamHI-BamHI frag- ments, as well as and adjacent 1.6 and 0.6 kB AvaI-AvaI fragments. This served to remove most or all of the coding regions of the idi (ORF6), crtX, crtY, and crtZ genes.
? Vector type
low copy number bacterial cloning vector
? Bacterial Resistance(s)
Chloramphenicol
? 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 lycopene production, or grow on agar plates at room temperature for 3-7 days. Use to confirm the function of the products of prospective DXS or IPI or LytB-encoding genes or cDNAs. Active enzymes will yield E. coli colonies much darker in color. Use pAtipiTrc or pdxs/ipiTrc as a positive control.
? Copy number
Unknown
? Gene/Insert name
crtE, crtI, crtB
? Species
Erwinia herbicola Eho10
? GenBank ID
M87280.1
? Promoterendogenous promoters

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.



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