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Expression of Myc-R9-EGFP fusion protein and validation of its transduction activity / 生物医学工程学杂志
Article em Zh | WPRIM | ID: wpr-271743
Biblioteca responsável: WPRO
ABSTRACT
To construct, express, purify and identify the Myc-R9-EGFP fusion protein and validate its transduction activity in the cultured porcine embryo fibroblasts. cDNA of pig c-Myc gene was amplified by RT-PCR with specific primers of 9 arginine (R9) from the primordial genital ridges and inserted into prokaryotic expression vector pET-28a-EGFP. After DNA sequencing confirmation, the recombinant plasmid was then transformed into BL21 (Escherichia coli) strain. After IPTG induction, the target fusion protein was efficiently induced to express, successfully purified by Novagen His-Bind kit, identified by SDS-PAGE and Western blotting. Finally, its high transduction activity in the porcine embryo fibroblasts was validated. The purified Myc-R9-EGFP fusion protein and the validation of its transduction activity in fibroblasts have provided an experimental foundation for further studies on the biological characterization of Myc protein, and soundly facilitated the further study of establishing pig induced pluripotent stem cells by recombinant protein.
Assuntos
Texto completo: 1 Índice: WPRIM Assunto principal: Arginina / Suínos / Proteínas Recombinantes de Fusão / Farmacocinética / Linhagem Celular / Permeabilidade da Membrana Celular / Proteínas Proto-Oncogênicas c-myc / Biologia Celular / Proteínas de Fluorescência Verde / Escherichia coli Limite: Animals Idioma: Zh Revista: Journal of Biomedical Engineering Ano de publicação: 2012 Tipo de documento: Article
Texto completo: 1 Índice: WPRIM Assunto principal: Arginina / Suínos / Proteínas Recombinantes de Fusão / Farmacocinética / Linhagem Celular / Permeabilidade da Membrana Celular / Proteínas Proto-Oncogênicas c-myc / Biologia Celular / Proteínas de Fluorescência Verde / Escherichia coli Limite: Animals Idioma: Zh Revista: Journal of Biomedical Engineering Ano de publicação: 2012 Tipo de documento: Article