|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||42922||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 3956
- Total vector size (bp) 9143
Vector typeWorm Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Bacterial Resistance(s)Ampicillin and Kanamycin
Growth Strain(s)ccdB Survival
Gene/Insert nameCe-age-1 promoter (873 bp)::Ce-age-1 cDNA (3549 bp)::Ce-unc-54 terminator (765 bp)
SpeciesC. elegans (nematode)
Insert Size (bp)5187
GenBank IDNM_064061.4 NP_496462.2
Entrez Geneage-1 (a.k.a. CELE_B0334.8)
- Promoter Ce-age-1 promoter (873 bp)
- Cloning method TOPO Cloning
- 5′ sequencing primer M13_pUC_fwd; T7
- 3′ sequencing primer M13_pUC_rev; T3 (Common Sequencing Primers)
The construct Ce-age-1p::Ce-age-1(3,549bp)::Ce-unc-54t, contained in plasmid pPV454, was assembled by overlap extension PCR. An 873 bp segment of Ce-age-1 5′ region was amplified from genomic DNA and fused to 2,376 bp of the 5′ region of Ce-age-1, amplified from cDNA, using the overlapping primers Ceage1p-cDNA-F and -R (below). This was subsequently fused to the remainder of the Ce-age-1 cDNA with the plasmid Pdpy-30-age-1, a gift from Yuichi Iino (University of Tokyo, Tokyo, Japan), as template and using the overlapping primers Ceage1-Cat-F and -R (below). Finally, the promoter and full-length 3,549 bp Ce-age-1 cDNA were fused to 765 bp of the Ce-unc-54 3′ region (terminator), amplified from Pdpy-30-age-1 template, using the overlapping primers Ceage1-unc54-F and -R (below). The PCR product was T/A cloned into the pCR4-TOPO vector (Life Technologies) and the insert fully sequenced.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pPV454 was a gift from James Lok (Addgene plasmid # 42922 ; http://n2t.net/addgene:42922 ; RRID:Addgene_42922)
For your References section:Strongyloides stercoralis age-1: a potential regulator of infective larval development in a parasitic nematode. Stoltzfus JD, Massey HC Jr, Nolan TJ, Griffith SD, Lok JB. PLoS One. 2012;7(6):e38587. doi: 10.1371/journal.pone.0038587. Epub 2012 Jun 6. 10.1371/journal.pone.0038587 PubMed 22701676