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为便于下游中试规模生产，采用温度敏感型启动子PRPL构建rhTNFα工程菌，并对影响工程菌发酵培养和表达的因素进行了初步研究。结果显示，采用RTB培养基，50℃热水快速升温，诱导培养4～4．5h，湿菌体收获率和表达量均达到较高水平；50L发酵罐连续发酵3批，14000r／min连续离心，菌体收获率湿重达16．gg／L培养物，rhTNFα表达量占菌体总蛋白的10.5％，活性达1．35×107U／mg蛋白。.
In order to facilitate downstream pilot-scale production, a temperature-sensitive promoter PRPL was used to construct rhTNFα-engineering bacteria, and preliminary studies were conducted on the factors affecting the fermentation and expression of the engineering bacteria. The results showed that using RTB medium, rapid heating with 50°C hot water, induction of culture for 4 to 4.5 hours, wet cell harvest rate and expression level reached a higher level; 50L fermentation tank continuous fermentation 3 batches, 14000r/min continuous centrifugation The rate of harvest of the bacteria reached a wet weight of 16. In gg/L cultures, the expression of rhTNFα accounted for 10.5% of total bacterial proteins, and the activity reached 1.35×107 U/mg protein. .