pAtipiTrc | Addgene 53275 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 :

pAtipiTrc

catalog :

53275

citations: 2

Reference
Berthelot K, Estevez Y, Quiliano M, Baldera Aguayo P, Zimic M, Pribat A, et al. HbIDI, SlIDI and EcIDI: A comparative study of isopentenyl diphosphate isomerase activity and structure. Biochimie. 2016;127:133-43 pubmed publisher
Cunningham F, Lafond T, Gantt E. Evidence of a role for LytB in the nonmevalonate pathway of isoprenoid biosynthesis. J Bacteriol. 2000;182:5841-8 pubmed

product information

Catalog Number :

53275

Product Name :

pAtipiTrc

article :

doi
id	8299
pubmed_id	11004185

bacterial resistance :

Ampicillin

cloning :

backbone

pBluescript SK-

backbone_mutation

A 0.85 kB KpnI-EcoRI fragment containing most of an Arabidopsis ipi1 cDNA [Cunningham, FX Jr, Gantt E (2000) Identification of Multi-Gene Families Encoding Isopentenyl Diphosphate Isomerase in Plants by Heterologous Complementation in Escherichia coli. Plant Cell Physiol. 41, 119-123.] was excised from the original cDNA library plasmid and ligated in the corresponding sites of plasmid pTrcHis B. Subsequent digestion with NcoI and XhoI, filling-in with the Klenow fragment of DNA polymerase, and ligation to recircularize, yielded a plasmid in which the coding region of ipi1, lacking the first 48 codons, is fused in frame to the strong bacterial Trc promoter and 6 additional codons that specify the amino acids MSRSAA. An EcoRV-EcoRI fragment, containing the cDNA together with the Trc promoter, was excised, gel-purified, and ligated in the corresponding sites of pBluescript SK to produce the plasmid pAtipiTrc (also referred to as clone 11-2).

backbone_origin

Stratagene

backbone_size

2958

promoter

sequencing_primer_3

sequencing_primer_5

vector_types

Bacterial Expression

growth strain :

Contains an Arabidopsis thaliana idi1 cDNA fused to a strong Trc promoter. Produces an active isopentenyl diphosphate isomerase (IDI) in E. coli.

growth temp :

This plasmid produces an active isopentenyl diphosphate isomerase (IPI) enzyme in E. coli, and can be used to enhance carotenoid production in E. coli containing carotenoid-producing plasmids, such as pAC-LYC and pAC-BETA and pAC-ZEAX, that lack a gene encoding an IPI. Use as a positive control when examining the activity of the products of prospective IPI-encoding genes or cDNAs in E. coli strain TOP10 (most other E. coli strains also are suitable) containing pAC-LYC or pAC-BETA. In this case, grow liquid cultures on a platform shaker at 28 degrees Celsius for 2-3 days in darkness, or grow on agar plates at room temperature for 3-7 days. An active IPI enzyme will result in E. coli colonies much darker in color.

origin :

pi :

alt_names

Atdyi

ipiAt1

cloning

clone_method	Restriction Enzyme
cloning_site_3	EcoRI
cloning_site_5	EcoRV
promoter	Trc
sequencing_primer_3	none
sequencing_primer_5	none
site_3_destroyed
site_5_destroyed

entrez_gene

aliases	AT3G02780, ATISOPENTENYL DIPHOSPHE ISOMERASE 2, F13E7.28, F13E7_28, IDI2, IPIAT1, ISOPENTENYL PYROPHOSPHATE:DIMETHYLLALLYL PYROPHOSPHATE ISOMERASE, isopentenyl pyrophosphate:dimethylallyl pyrophosphate isomerase 2
gene	IPP2
id	820960

genbank_ids

AF188066.1

mutation

name

ipi1

shRNA_sequence

size

1595

species

3702

Arabidopsis thaliana