| 姚占娟，张其中，崔淼.斜带石斑鱼IgZ重链基因的表达纯化和抗体制备[J].广东农业科学,2013,40(5):149-152 |
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斜带石斑鱼IgZ重链基因的表达纯化和抗体制备 |
| Prokaryotic expression, purification of orange-spotted grouper(Epinephelus coioides) IgZ heavy-chain protein and preparation of polyclonal antibody against IgZ |
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| DOI： |
| 中文关键词: 斜带石斑鱼 免疫球蛋白Z 原核表达 多克隆抗体制备 |
| 英文关键词: Epinephelus coioides immunoglobulins Z (IgZ) prokaryotic expression antibody preparation |
| 基金项目:国家自然科学基金广东省人民政府联合项目（U0631010）；国家自然科学基金（40576056，40976066，31170474）；广东省自然科学基金（04300664，07300378） |
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| 中文摘要: |
| 为获得斜带石斑鱼IgZ 重链基因的多克隆抗体，成功构建了斜带石斑鱼免疫球蛋白IgZ 重链重组蛋白表达质粒pQE30/IgZ，将其转化到大肠杆菌表达菌株M15 中，确定了最佳诱导表达条件：IPTG 0.2 mmol/L，诱导温度30℃，诱导时间6 h。所获得的IgZ 重链重组蛋白经Ni-NTA 亲和层析后，纯度达到85%以上；以纯化的IgZ重链重组蛋白为抗原免疫新西兰兔，制备出相应的多克隆抗体，经ELISA 测定抗血清效价约为1：320 000。 |
| 英文摘要: |
| 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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