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KMID : 0364519950070020151
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Dong-A Journal Medicine
1995 Volume.7 No. 2 p.151 ~ p.160
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Expression of rat liver nucleolar phosphoprotein B23 in Escherichia coli
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Abstract
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phoprotein B23 is one of the major protein components in eukaryotic nucleolus. The cellular content and distribution of this protein are correlated with the synthetic rate of ribosomal RNA and cell growth, It was attempted to produce this protein
in
Escherichia coli, using pGEMEX vector system based on bacteriophage T7 RNA polymerase. Subcloning of B23 cDNA into pGEMEX vector was performed by ligating the fragment to EcoRI and BamHI sites. Adding IPTG at the 0.5mM concentration, protein
synthesis
was induced. After electrophoresis of bacterial crude extracts in SDS-PAGE, expression of B23 cDNA was analyzed by anti-B23 monoclonal antibody. Immunoblot shows that subcloned gene was successfully expressed as 80 kD fusion protein with leader
sequences of T7 gene 10 in Escherichia coli. From these results, we concluded that eukaryotic B23 gene could be expressed in prokaryotic cells with its intact epitope structure.
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KEYWORD
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