Expression and Purification of Recombinant Mouse Ets-1 Transcription Factor

• Published in: Protein expression and purification Vol. 21; no. 3; pp. 492 - 499
• Main Authors: Lionneton, Frédéric, Drobecq, Hervé, Soncin, Fabrice
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-04-2001; Elsevier
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cadherins - genetics; Cardiology and cardiovascular system; Chromatography, Affinity; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; DNA Probes - genetics; DNA Probes - metabolism; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; DNA-Binding Proteins - isolation & purification; DNA-Binding Proteins - metabolism; Electrophoresis, Polyacrylamide Gel; Genetic Vectors; Human health and pathology; Life Sciences; Mass Spectrometry; Mice; Molecular Sequence Data; Promoter Regions, Genetic - genetics; Proto-Oncogene Protein c-ets-1; Proto-Oncogene Proteins - chemistry; Proto-Oncogene Proteins - genetics; Proto-Oncogene Proteins - isolation & purification; Proto-Oncogene Proteins - metabolism; Proto-Oncogene Proteins c-ets; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Response Elements - genetics; Transcription Factors - chemistry; Transcription Factors - genetics; Transcription Factors - isolation & purification; Transcription Factors - metabolism
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1006/prep.2001.1405

Ets-1 is a transcription factor which belongs to the ETS family. Its mRNA is expressed in the embryo during normal development and also in tumors. In order to sort out functional Ets-1-binding sites among those present in gene promoters, we constructed an expression vector and designed a purification protocol for the production of the 440-amino-acid form of mouse Ets-1, based on heparin-Sepharose affinity and anion-exchange chromatographies. This protocol allows the purification of large amounts of pure recombinant protein as assessed by SDS-PAGE, C18 reverse-phase HPLC, amino-terminal sequencing, and mass spectrometry. The purified protein is recognized by specific anti-Ets-1 antibodies and binds to DNA ETS-binding sites.