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| Prokaryotic Expression, Analysis and Identification of Human Polypyrimidine Tract Binding Protein-associated Splicing Factor |
| WANG Hui-juan1, LAN Yang1, LIU Tao1, LI Ling1,2 |
1. Functional Genome Laboratory, College of Life Science, Sichuan University, Chengdu 610064, China;
2. Hainan Provincial Key Laboratory of Tropical Medicine, Hainan Medical School, Haikou 571101, China |
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Abstract Polypyrimidine tract binding protein-associated splicing factor (PSF), a multifunctional RNA/DNA-binding protein, can suppress the expression of proto-oncogene and function as tumor suppressor protein (TSP) in human and mouse. Using total RNA extracted from human fibroblasts, PSF-encoding cDNA was synthesized and inserted into vector pET-28a(+) to construct the recombinant plasmid pET-28-PSF. The PSF fused with His-tag was expressed in E.coli BL21 (DE3) by IPTG induction and then purified by affinity chromatography. SDS-PAGE and Western blotting results indicated the soluble His-tagged PSF could be produced with a prokaryotic expression system. Finally, the RNA-binding capacity of recombinant PSF to mVL30-1 RNA was determined by Gel-shift assay. The results indicate that recombinant PSF produced with a prokaryotic expression system can be biologically active and will therefore allow us to investigate the function of PSF in the future.
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Received: 21 February 2011
Published: 28 June 2011
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Corresponding Authors:
Li Ling
E-mail: leolibeike@gmail.com
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