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文档编号:SW078 文档字数:11888,页数:22 摘 要：本文研究重组菌plyC高密度发酵在大肠杆菌中诱导表达的条件。首先利用自控式5L发酵罐进行链球菌噬菌体裂解酶A链（plyCA）工程菌补料式发酵，对补料时机、诱导时间、诱导剂量对链球菌噬菌体裂解酶(plyC)A链和B链（plyCA,plyCB）在大肠杆菌中表达情况的影响进行研究，以SDS-PAGE检测方法来检测目的蛋白表达情况。结果表明，补料时机在发酵进行3.5 h后加入，以一定速度连续加料，并加入诱导剂培养。另以1.7 mmoL/L IPTG诱导表达实验表明，诱导时间为4小时链球菌噬菌体裂解酶重组菌表达的目的产物量最高。用30 µL IPTG诱导使其终浓度为1 mmoL/L可使PLYCA和PLYCB的表达量相对较高。 Abstract：This article to investigate the optimal high-density fermentation of recombinant bacteria plyC induced in E. coli conditions. Firstly, make use of 5L autocontrol fermentor for streptococcal bacteriophage lysin chain A (plyCA) fed batch fermentation bacteria feeding on feeding time, induced time, induced dose of the streptococcus bacteriophage lyase (plyC) A chain and B chain ( plyCA, plyCB) expressed in E. coli to study the impact of the situation to SDS-PAGE detection method to detect protein expression. The results show that the feeding time after 3.5 h in the fermentation by adding, to a certain speed continuous feeding, and the addition of induction agent training. The other to 1.7 mmol/L IPTG induced expression experiments show that the induction time of 4 hours lyase of streptococcal bacteriophage lysin recombinant expression produced of the highest purpose. Induced with 30 μL IPTG to a final concentration of 1 mmoL/L may cause PlyCA and the PlyCB expression quantity is relatively high
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| Tags：链球菌 裂解 大肠杆菌 表达 初步 研究 | 2011-03-31 15:42:40【返回顶部】 |
