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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
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>
Subject(s)
Full text: 1 Index: WPRIM Main subject: Pathology / Prostatic Neoplasms / Recombinant Proteins / Molecular Sequence Data / Base Sequence / Transfection / Gene Expression / Retinoblastoma Protein / Cloning, Molecular / Cell Line, Tumor Limits: Humans / Male Language: Zh Journal: Journal of Southern Medical University Year: 2010 Type: Article
Full text: 1 Index: WPRIM Main subject: Pathology / Prostatic Neoplasms / Recombinant Proteins / Molecular Sequence Data / Base Sequence / Transfection / Gene Expression / Retinoblastoma Protein / Cloning, Molecular / Cell Line, Tumor Limits: Humans / Male Language: Zh Journal: Journal of Southern Medical University Year: 2010 Type: Article