PMID: 15367352Sep 16, 2004Paper

Expression of GST-PRL-2 fusion protein in prokaryotic cells and preparation of Hen egg yolk immunoglobulin (IgY) against PRL-2

Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Ai-Lin GuoJian-ming Wen

Abstract

To express phosphatase 2 of regenerating liver (PRL-2) fusion protein in E.coli and to prepare specific hen egg yolk immunoglobulin(IgY) against PRL-2. A full-length human PRL-2 gene coding sequence was cloned into expression vectors pGEX-4T-2 and pET21a, then transformed into E.coli. Fusion protein GST-PRL-2 was expressed in E.coli via IPTG induction. The expressed proteins were purified from lysates through Glutathione Sepharose 4B and the Ni-NTA agarose columns, respectively. Egg-laying hens were immunized using the purified GST-PRL-2 with Freund's complete or incomplete adjuvant. The specificity of the resulting antibody was identified by Western blot. A high level of expression of target protein was detected by Western blot after IPTG induction and purified protein was obtained through Glutathione Sepharose 4B and the Ni-NTA affinity chromatography agarose columns, respectively. Western blot analysis showed that the anti-PRL-2 polyclonal antibody can recognize 6 x His-PRL-2 fusion protein. The hen egg yolk immunoglobulin(IgY) against PRL-2 expressed in E.coli has good specificity which provides an useful reagent for the detection of PRL-2.

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