[Article in Chinese]
Guo AL, Cao YX, Wang W, Ding B, Huang YQ, Wen JM. Department of Pathology, Zhongshan Medical College, SUN Yat-sen University, Guangzhou 510080,China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 Sep;20(5):578-81.
Abstract
AIM: 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.
METHODS: 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.
RESULTS: 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 6xHis-PRL-2 fusion protein.
CONCLUSION: 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. PMID: 15367352 [PubMed - in process] | 
