PurposeExpression of luciferase reporter mRNA in S2 cell
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||50555||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerThe Drosophila Gateway™ Vector Collection
Vector typeInsect Expression, Luciferase
Growth in Bacteria
- Promoter Actin5C
- Cloning method Gateway Cloning
- 5′ sequencing primer RenLuc3'F (Common Sequencing Primers)
We constructed a Renilla luciferase reporter for let-7 miRNA harboring A114-N40 (a protected poly(A) sequence internalized by a 40 nt unrelated sequence) followed by the hammerhead ribozyme sequence (HhR) that generates the 3' end by self-cleavage.
Unstable when transformed and so any retransformation must be checked by digest or sequencing before use. In strain MV1184, it is stable as a glycerol stock.
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:pAWH-Rluc-let-7-A114-N40-HhR was a gift from Yukihide Tomari (Addgene plasmid # 50555 ; http://n2t.net/addgene:50555 ; RRID:Addgene_50555)
For your References section:MicroRNAs mediate gene silencing via multiple different pathways in drosophila. Fukaya T, Tomari Y. Mol Cell. 2012 Dec 28;48(6):825-36. doi: 10.1016/j.molcel.2012.09.024. Epub 2012 Nov 1. 10.1016/j.molcel.2012.09.024 PubMed 23123195