Purposemammalian expression of GFP-tagged human centromere protein A
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||117803||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector typeMammalian Expression
Growth in Bacteria
Copy numberHigh Copy
SpeciesH. sapiens (human)
Insert Size (bp)430
MutationSee depositor comments below.
Entrez GeneCENPA (a.k.a. CENP-A, CenH3)
/ Fusion Protein
- GFP (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site SacI (not destroyed)
- 3′ cloning site XhoI/SalI (destroyed during cloning)
- 5′ sequencing primer CMV-F (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
The insert fragment with Sac I and Xho I sites was cloned into Sac I and Sal I sites of pEGFP-C1.
Thus, there is no Xho I and Sal I site in pEGFP-AF8.
The stop codon was also changed to TCG (..ctcggcTCGac-3’)
Note that the cloning strategy adds additional amino acids onto the C-terminus of the insert (STVPRARDPPDLDN), but this does not interfere with the function.
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:pEGFP-AF8 was a gift from Kenji Sugimoto (Addgene plasmid # 117803 ; http://n2t.net/addgene:117803 ; RRID:Addgene_117803)
For your References section:Centromere/kinetochore localization of human centromere protein A (CENP-A) exogenously expressed as a fusion to green fluorescent protein. Sugimoto K, Fukuda R, Himeno M. Cell Struct Funct. 2000 Aug;25(4):253-61. PubMed 11129795