| 摘要: |
| [目的]探讨原核表达的柔嫩艾美耳球虫杨凌株3-1E蛋白的生物学活性。[方法] 将含有3-1E基因的pGEX-3-1E重组质粒转化大肠杆菌BL21,进行表达并对表达条件进行优化,表达产物纯化后进行Western-blotting分析。[结果] 表达的3-1E重组蛋白相对分子质量为44。7 ku,与推导结果一致。在28 ℃、IPTG诱导浓度为0。5 mmol/L的条件下诱导6 h,可溶性3-1E重组蛋白表达量最高。Western-blotting结果表明,3-1E重组蛋白可与兔抗鸡E。tenella YL阳性血清发生反应。[结论] 3-1E重组蛋白表达成功,表达产物具有免疫原性。 |
| 关键词: 3-1E基因 柔嫩艾美耳球虫 基因表达 生物活性分析 |
| DOI： |
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| 基金项目:国家“863”计划项目（2006AA10A207） |
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| Expression of E.tenella YL 3-1E gene and bioactivity analysis of the production |
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| Abstract: |
| 【Objective】 The paper investigated bioactivity of Eimeria tenella 3-1E protein expressed in E.coli.【Method】 The recombinant plasmid pGEX-3-1E sequenced correctly was transformed into E.coli BL21,and was prokaryotic expressed in E.coli.After optimizing expression conditions,the purified proteins were Western-Blotted.【Result】 The expression fusion protein was a 44.7 ku,and was in agreement with deduction,the highest soluble expression level of recombinant protein was obtained after being induced at 28 ℃ for 6 h and the concentration of IPTG was 0.5 mmol/L.The purified proteins could react with the anti-sera derived from rabbits challenged with the E.tenella YL strain.【Conclusion】 The 3-1E protein possesses immunogenicity. |
| Key words: 3-1Egene Eimeria tenella gene expression bioactivity analysis |
