Construction, expression and location of eukaryotic expression vector CMV-FLAG-RB in human prostate cancer PC-3 cells / 南方医科大学学报
Journal of Southern Medical University
; (12): 422-425, 2010.
Article
in Zh
| WPRIM
| ID: wpr-269538
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct an eukaryotic recombinant expression vector for retinoblastoma 1 gene (RB-1) and investigate the role of RB-1 in prostate cancer.</p><p><b>METHODS</b>The coding sequence of RB-1 gene tagged with FLAG was amplified from the plasmid CMV-RB by PCR method. The fragment was cloned into CMV expression vector and identified by restriction enzyme digestion and sequence analysis. Western Blotting was used to detect RB-1 expression and immunofluorescence was used to observe RB-1 distribution in PC-3 cells transfected with the recombinant.</p><p><b>RESULTS</b>The expression vector CMV-FLAG-RB was successfully constructed as confirmed by PCR, endonuclease digestion and DNA sequence analysis. RB-1 protein was highly expressed and showed a nuclear distribution in PC-3 cells transfected with the recombinant.</p><p><b>CONCLUSIONS</b>The eukaryotic expression vector for RB-1 has been successfully constructed and can be efficiently expressed in PC-3 cells. The expression of RB-1 is located in the cell nuclei.</p>
Full text:
1
Index:
WPRIM
Main subject:
Pathology
/
Prostatic Neoplasms
/
Recombinant Proteins
/
Molecular Sequence Data
/
Base Sequence
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Transfection
/
Gene Expression
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Retinoblastoma Protein
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Cloning, Molecular
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Cell Line, Tumor
Limits:
Humans
/
Male
Language:
Zh
Journal:
Journal of Southern Medical University
Year:
2010
Type:
Article