Aim: To express and purify Lugurus zone pellucida 3 (LZP3) in prokaryotic cells and to prepare the LZP3-specific rabbit antiserum.
Methods: The core fragment of LZP3 gene was cloned into plasmid pGEX-4T-1 containing glutathione s-transferase (GST) fusion protein gene. Following restriction enzyme digestion analysis and sequencing, pGEX-4T-LZP3 was transformed into E. coli BL21(DE3). GST-LZP3 fusion protein was expressed under IPTG induction and further purified with Glutathione Sepharose 4B. Then the purified GST-LZP3 fusion protein was used to immunize New Zealand rabbits. LZP3-specific rabbit antiserum was identified by ELISA and Western blot.
Results: GST-LZP3 fusion protein was overexpressed and its LZP3-specific antiserum was obtained.
Conclusion: The successful expression of GST-LZP3 fusion protein in E.coli and the preparation of LZP3-specific rabbit antiserum will be valuable for the study on birth control of Lagurus.