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| Prokaryotic expression, purification of orange-spotted grouper(Epinephelus coioides) IgZ heavy-chain protein and preparation of polyclonal antibody against IgZ |
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| Abstract: |
| The recombinant expression plasmid was constructed by means of linking the grouper IgZ cDNA with prokaryotic
expression vector pQE30. It was identified by endonuclease digestion and DNA sequencing of the recombinant plasmid. Then, the recombinant plasmid pQE30/IgZ was transformed into E. coli M15. And the most optimum induced expression conditions were determined:the IPTG was 0.2 mmol/L, the induced temperature was 30益, the induction time was 6 h. The recombinant IgZ heavy-chain protein was gained by means of Ni-affinity chromatography. The purity of the recombinant IgZ heavy-chain protein was more than 85%. The anti-IgZ polyclonal antibody was gotten by means of immuning New Zealand white rabbit with the recombinant IgZ heavy chain protein. The result of Enzyme-linked Immunosorbent Assay (ELISA) showed that the highest titer of anti-serum was 1：320 000 for IgZ. |
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