pAC-DELTA | Addgene 53273 product information

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product summary

company name :

Addgene

product type :

cDNA

product name :

pAC-DELTA

catalog :

53273

citations: 2

Reference
Loewen P, Switala J, Wells J, Huang F, Zara A, Allingham J, et al. Structure and function of a lignostilbene-α,β-dioxygenase orthologue from Pseudomonas brassicacearum. BMC Biochem. 2018;19:8 pubmed publisher
Sun Z, Gantt E, Cunningham F. Cloning and functional analysis of the beta-carotene hydroxylase of Arabidopsis thaliana. J Biol Chem. 1996;271:24349-52 pubmed

product information

Catalog Number :

53273

Product Name :

pAC-DELTA

article :

doi
id	8298
pubmed_id	8798688

bacterial resistance :

Chloramphenicol

cloning :

backbone

pAC-LYC

backbone_mutation

A 1.9-kb SmaI-EcoRV fragment containing an A. thaliana lycopene cyclase cDNA was excised from plasmid y2 and cloned in the Klenow-blunted BamHI site of pTrcHis A to give pTrc-At . A 2.5-kb EcoRV-KpnI fragment containing the Trc promoter and the A. thaliana lycopene cyclase was then excised from pTrc-At and cloned in the Klenow-blunted HindIII site of pAC-LYC to produce pAC-DELTA. The reasoning behind this cloning strategy is now obscure, but the resulting plasmid is useful, yielding delta-carotene as the predominant carotenoid in E. coli. The orientation of the lycopene epsilon-cyclase cDNA relative to the Trc promoter, or to the p15A origin of replication for this plasmid, are both uncertain, therefore the sequence submitted for this plasmid may not be correct.

backbone_origin

Francis X. Cunningham, Jr.

backbone_size

9476

promoter

sequencing_primer_3

sequencing_primer_5

vector_types

Other

low copy number bacterial cloning vector

growth notes :

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.

growth strain :

Contains crtE, crtB, and crtI genes of Erwinia herbicola (Pantoea agglomerans) Eho10, together with the lcyE gene of Arabidopsis thaliana and thereby produces delta-carotene in E. coli

growth temp :

Grow liquid cultures on a platform shaker at 28 degrees Celsius for 2-3 days in darkness for best delta-carotene production, or grow on agar plates at room temperature for 3-7 days.

origin :

pi :

alt_names

lycopene epsilon cyclase

lycopene epsilon-monocyclase

cloning

clone_method	Restriction Enzyme
cloning_site_3	HindIII
cloning_site_5	HindIII
promoter	Trc
sequencing_primer_3	none
sequencing_primer_5	none
site_3_destroyed	1
site_5_destroyed	1

entrez_gene

aliases	AT5G57030, LUTEIN DEFICIENT 2, LYCOPENE EPSILON-CYCLASE
gene	LUT2
id	835806

genbank_ids

U50738.1

mutation

name

lcyE

shRNA_sequence

size

species

3702

Arabidopsis thaliana