Recombinant prokaryotic plasmid construction and high expression of FUS1 gene / 南方医科大学学报
Journal of Southern Medical University
;
(12): 638-640, 2007.
Article
in Chinese
| WPRIM
| ID: wpr-268060
ABSTRACT
<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>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Transformation, Genetic
/
Umbilical Cord
/
Recombinant Proteins
/
Blotting, Western
/
Cloning, Molecular
/
Cell Biology
/
Tumor Suppressor Proteins
/
Electrophoresis, Polyacrylamide Gel
/
Escherichia coli
Limits:
Humans
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2007
Type:
Article
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