| 摘要: |
| 以重组别藻蓝蛋白表达工程菌JM109(DE3)/pET28-APC 作为研究对象，对于JM109(DE3)菌株在乳糖诱导下表达重组蛋白的规律进行了研究。比较分析了不同生长阶段进行诱导，最佳乳糖浓度、诱导持续时间和诱导温度等参数对重组蛋白表达的影响。实验结果表明，采用JM109(DE3)为宿主菌，经过条件优化，乳糖诱导重组蛋白的表达量可以达到IPTG 诱导的水平；较低的诱导培养温度能有效提高可溶性重组蛋白的表达。最后在摇瓶发酵结果的基础上，实现了乳糖诱导工程菌的5L 全自动发酵罐培养。研究结果为在大肠杆菌大规模发酵中，运用廉价、无毒的乳糖作为诱导剂提供了一定的借鉴。 |
| 关键词: 乳糖 低温诱导 别藻蓝蛋白 大肠杆菌 |
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| 基金项目:国家863 计划项目（2001AA620410）;中国科学院创新工程资助项目（KZCX3–SW–215） |
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| Lactose-induced expression of recombinant allophycocyanin in Escherichia coli JM109(DE3) |
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| Abstract: |
| The expression recombinant allophycocyanin gene in Escherichia coli JM109(DE3) using lactose as the inducer was optimized. The influences of culture conditions, including the point of induction, the lactose concentration, the duration of induction phase and the induction temperature were analyzed. Finally the recombinant E. coli was cultured in a 5L fermentor. The result showed that lactose could be used to induce recombinant allophycocyanin expression to concentrations exceeding the level achieved by using with IPTG as an inducer. After adding lactose, the low culture temperature could increase the expression level of soluble target protein. |
| Key words: low-temperature induction Allophycocyanin Escherichia coli |
