Preparation and characterization of a monoclonal antibody specific to histone acetyltransferase from Plasmodium falciparum.
We have obtained a specific monoclonal antibody (MAb) against the Plasmodium falciparum histone acetyl transferase (PfGcn5), a transcriptional factor that possesses HAT activity directed to the amino terminal of histone H3. To prepare this antibody, a 968-base pair (bp) DNA fragment of PfGcn5 gene corresponding to C-terminal domain was obtained by RT-PCR and cloned into prokaryotic expression vector pGEX-4T3. MAb against PfGcn5 was obtained with hybridoma technique and ELISA screening using either purified GSTPfGcn5 protein or purified GST protein alone as a control. One MAb, named Pf.r2, was able to identify the PfGcn5 protein in nuclear extract from P. falciparum and immunofluorescence assays. This MAb will be a helpful tool to perform a variety of assays to identify the other components of PfGcn5 complexes.
Montiel-Condado
Dvorak
D
Department of Molecular Biomedicine, Centro de Investigación y de Estudios Avanzados del IPN, D.F., México.
Romero-Ramírez
Héctor
H
Ramírez-Estudillo
Carmen
C
Santos-Argumedo
Leopoldo
L
Hernández-Rivas
Rosaura
R
D000123:Acetyltransferases / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D004912:Erythrocytes / Q000276:immunology / Q000469:parasitology; D051548:Histone Acetyltransferases; D006801:Humans; D006825:Hybridomas; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017423:Sequence Analysis, RNA
D000911:Antibodies, Monoclonal; D000123:Acetyltransferases; D051548:Histone Acetyltransferases