pAC-BETA-At | Addgene 53288 product information

This webpage contains legacy information. The product is either no longer available from the supplier or has been delisted at Labome.

product summary

company name :

Addgene

product type :

cDNA

product name :

pAC-BETA-At

catalog :

53288

citations: 2

Reference
Gupta P, Rodriguez Franco M, Bodanapu R, Sreelakshmi Y, Sharma R. Phytoene synthase 2 in tomato fruits remains functional and contributes to abscisic acid formation. Plant Sci. 2022;316:111177 pubmed publisher
Cunningham F, Lee H, Gantt E. Carotenoid biosynthesis in the primitive red alga Cyanidioschyzon merolae. Eukaryot Cell. 2007;6:533-45 pubmed

product information

Catalog Number :

53288

Product Name :

pAC-BETA-At

article :

doi	10.1128/EC.00265-06
id	8301
pubmed_id	17085635

bacterial resistance :

Chloramphenicol

cloning :

backbone

pAC-LYC

backbone_mutation

The entire open reading frame of an Arabidopsis lcyB cDNA [GenBank U50739.1] in plasmid pZL1 was excised as an EcoRI-HindIII fragment of ca. 1.8 kB and cloned in the same sites of pBluescript SK- so that the cDNA was fused, in frame, to the N terminal portion of the lacZ gene and under the control of the lac promoter. From this plasmid (pAtLCYbSK) a ca. 1.8-kB NotI-NotI fragment containing the lcyB cDNA (without the lac promoter) was excised, made blunt with the Klenow enzyme, and cloned in the Klenow-blunted HindIII site of pAC-LYC. The lcyB cDNA in this 11.3 kB plasmid was ascertained, by restriction enzyme digestion, to be in the forward orientation. Despite the lack of a bacterial promoter, production of the LCYb enzyme is sufficient so that beta-carotene is the major carotenoid in E. coli, but also low enough so that the introduction of a second plasmid with a strongly-expressed lycopene epsilon cyclase cDNA (i.e plasmid pY2F) will yield predominantly alpha-carotene in E. coli.

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, and an lcyB cDNA of Arabidopsis thaliana. Use with pY2F to produce alpha-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 beta-carotene production, or grow on agar plates at room temperature for 3-7 days. Combine with plasmid pY2F and grow liquid cultures on a platform shaker at 28 degrees Celsius for 2-3 days in darkness for best alpha-carotene production, or grow on agar plates at room temperature for 3-7 days.

origin :

pi :

alt_names

lycopene beta-cyclase

cloning

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

entrez_gene

aliases	AT3G10230, LYCOPENE BETA-CYCLASE, lycopene cyclase
gene	LYC
id	820185

genbank_ids

U50739.1

mutation

name

lcyB

shRNA_sequence

size

1824

species

3702

Arabidopsis thaliana