TOPO-FLAG-GATA4(M456)
| Backbone | pCR-BluntII-TOPO |
|---|---|
| Insert | FLAG-mGATA4(K311A/K318A/K320A/K322A) |
| Seq | cDNA / amino acid |
| Site | EcoRI/XhoI |
| Selection | Kan |
Insertion
TOPO...M13 Fwd→...EcoRI-EcoRI-FLAG-GATA4(M456)-XhoI→-EcoRI...←M13 Rev...TOPO
Note
K311, K318, K320, and K322 of GATA4 are acetylated by p300. GATA4(M456) is lacking p300-induced acetylation, DNA-binding, and transcriptioanl activities. GATA4(M456) serves as a dominant-negative form because it can interact with its partners including p300 (Takaya, 2008).
FLAG-GATA4(M456) cDNA was amplified by using pcDNA-FLAG-GATA4(M456) as a template.
Reference
Takaya T, Kawamura T, Morimoto T, Ono K, Kita T, Shimatsu A, Hasegawa K. Identification of p300-targeted acetylated residues in GATA4 during hypertrophic responses in cardiac myocytes. J Biol Chem. 2008; 283: 9828-35.