Expression of recombinant mouse Raf-1 in escherichia coli
Dongchun Liang · Tianjin Yike Daxue xuebao · 2007
Objective: To express recombinant mouse Raf-1 in E.coli and make a basis for studying the role of Raf-1 in multiple cell signaling pathways. Methods: Total RNA were extracted from mouse liver,Raf-1 gene was amplified by a Nest-PCR following reverse transcription. The Raf-1 gene was cloned into plasmid pETDuet-1 by KpnI/NdeI digestion and the insertion was verified by DNA sequencing.Recombinant plasmid was transformed into E.coli DH5α and then subjected to 0.1mmol/L IPTG induced expression. Expression product was analyzed by SDS-PAGE and Western blot. Results:The recombinant mouse Raf-1 existed in the pellet of cell lysate. Conclusion:Recombinant mouse Raf-1 is expressed successfully in E.coli.