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Journal of Southern Medical University ; (12): 638-640, 2007.
Artigo em Chinês | WPRIM | ID: wpr-268060

RESUMO

<p><b>OBJECTIVE</b>To construct the prokaryotic plasmid of FUS1 gene for efficient FUS1 expression in E.coli strain Rosetta(DE3)2plys.</p><p><b>METHODS</b>The full-length FUS1 gene was amplified by PCR from the total RNA of umbilical mesenchymal stem cells and cloned into pET-32a(+) vector followed by identification with PCR and sequencing. The recombinant plasmid pET-32a(+)-FUS1 was transformed into the E.coli strain Rosetta(DE3)2plys and the target protein expression was induced by IPTG.</p><p><b>RESULTS</b>The plasmid pET-32a(+)-FUS1 was obtained successfully as verified by PCR and sequence analysis. High expression of the fused FUS1 protein was achieved after induction by low-concentration IPTG (25 micromol/L) for 3 h, and the recombinant FUS1 protein accounted for 40% of the total bacterial protein of Rosetta(DE3)2plys.</p><p><b>CONCLUSION</b>The recombinant FUS1 plasmid has been successfully cloned, which allows highly efficient FUS1 expression in Rosetta (DE3)2 plys.</p>


Assuntos
Humanos , Western Blotting , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Escherichia coli , Genética , Células-Tronco Mesenquimais , Biologia Celular , Metabolismo , Plasmídeos , Genética , Proteínas Recombinantes , Transformação Genética , Proteínas Supressoras de Tumor , Genética , Cordão Umbilical , Biologia Celular
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