【摘要】 采用PCR方法从pGEM-IL-18重组质粒中亚克隆出IL-18基因并完成了真核融合表达载体pPIC9K-IL-18的构建和IL-18在毕赤酵母中的表达,在此基础上从发酵时间、不同诱导型、pH值、诱导剂量和接种量等方面对重组基因工程菌的摇瓶发酵条件进行了优化,进一步探索了酵母菌表达猪IL-18的发酵工艺.结果表明,摇瓶发酵时,诱导最佳时间为72 h,甲醇最适浓度为20 g/L,最适pH范围为6.0~8.0,最适接种量1∶1.与单纯甲醇诱导相比,甘油/甲醇诱导后的表达量明显提高.更多还原

【Abstract】 The porcine IL-18 gene was amplified from recombinant plasmid pGEM-IL-18 by PCR,the positive expression vector,designated as pPIC9K-IL-18,was constructed and induced by methanol.Different culturing time,inducible pattern,pH and the amount of inducible reagents were compared,respectively.The results revealed that the optimal fermentation timing is was 72 h after induction with methanol,the optimal concentration of methanol was 20 g/L,the optimal pH range was 6.0～8.0 and the optimized inoculation amount was 1∶1.Compared with the induction only with methanol,combined use of glycerol and methanol could induce higher expression of the interest protein.更多还原