[Expression of Lagurus ZP3 fusion protein in prokaryotic cells and preparation of its antiserum].
OBJECTIVE
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.


CONCLUSIONS
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.
Zhang
Ai-lian
AL
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China. xjzal@163.com
Zhao
Gan
G
Wang
Bin
B
Zhang
Fu-chun
FC
D000818:Animals; D004527:Egg Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D004926:Escherichia coli / Q000378:metabolism*; D005260:Female; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics; D007106:Immune Sera / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D009115:Muridae; D010957:Plasmids; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D014170:Transformation, Genetic; D000072216:Zona Pellucida Glycoproteins
D004527:Egg Proteins; D007106:Immune Sera; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D000072216:Zona Pellucida Glycoproteins; D005982:Glutathione Transferase