| 摘要: |
| 【目的】获得鸡α干扰素重组酵母菌诱导表达的最佳诱导时间和甲醇的体积分数，并测定诱导表达产物对鸡马立克氏病毒（MDV）和鸡新城疫病毒（NDV）的抑制能力。【方法】 挑取2个鸡α干扰素重组酵母菌单菌落至BMGY培养基中培养，待菌液OD600为4左右时，分别转接至BMMY培养液诱导，1瓶每隔12 h补加1次体积分数0.5%甲醇，另1瓶每隔12 h补加1次体积分数1%甲醇，于24，48，72 h收集样品，备SDS-PAGE检测用。根据不同时间（24，48，72，96 h）诱导上清液表达量的差异，留用表达量最高时段的上清液，经初步纯化后，利用鸡胚成纤维细胞（CEF），分别测定重组酵母鸡α干扰素抑制MDV及NDV的能力。【结果】 鸡α干扰素重组酵母菌在每隔12 h补加1次体积分数1%甲醇诱导48 h时,或每隔12 h补加1次体积分数0.5%甲醇诱导72 h时, 重组酵母鸡α干扰素表达量均达到最高;且诱导上清液具有较好地抑制MDV、NDV的能力。【结论】 获得了鸡α干扰素重组酵母菌小量发酵的最佳条件；获得的重组酵母鸡α干扰素对MDV和NDV有明显的抑制作用。 |
| 关键词: 鸡α干扰素 酵母 MDV NDV |
| DOI： |
| 分类号: |
| 基金项目:江苏大学高级人才专项资助项目（1283000152） |
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| The fermenting optimization of the recombinant chicken interferon α yeast and detection for its antiviral activity to MDV and NDV |
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| Abstract: |
| 【Objective】The research was to find the optimal conditions for induction and volume fraction of the recombinant ChIFN-α yeast and study its activity of inhibiting Markers Disease virus (MDV) and Newcastle Disease virus (NDV).【Method】Two recombinants were induced at different time and different concentrations of methanol,the production of recombinant ChIFN-α was detected by SDS PAGE.The antivirus activity to MDV and NDV of the ChIFN-α was detected in the chicken embryonic fibroblast (CEF).【Result】 The optimal induction time was in the 48 h with 1%（V/V）methanol added into culture at every 12 h;the optimal induction time was in the 72 h with 0.5%（V/V）methanol added into culture at every 12 h and into culture.The recombinant chicken interferon expressed in yeast was of better activity of inhibitting MDV and NDV.【Conclusion】 The fermenting optimization of the recombinant chicken interferon-α has been obtained,which can inhibit the multiplication of MDV and NDV highly. |
| Key words: chicken alpha interferon yeast MDV NDV |
