诱导条件对重组大肠杆菌发酵中人白细胞介素6表达量的影响

INFLUENCE OF INDUCTION CONDITIONS ON THE EXPRESSION ON HUMAN INTERLEUKIN-6 IN RECOMBINANT E.coli

对重组人白细胞介素 6(rh IL- 6)大肠杆菌摇瓶发酵工艺作了研究。探讨了诱导策略对菌体生长和rh IL- 6表达量的影响。结果表明 ,含有温敏扩增型质粒 p Kp L3a的重组 h IL- 6大肠杆菌 ,在 2× YT培养基中发酵 ,接种量为 0 .2 % ,于 30℃ ,1 80 r/ min摇瓶中培养 ,当菌密度 OD60 0 约为 1 .5时 ,转至 42℃或 40℃诱导 ,维持时间至少为 1 h,然后转至 37℃培养 3h,最终 rh IL- 6表达量可达 32 %。于 40℃或 42℃下诱导时间低于1 h,则使最终 rh IL- 6表达量降低。如果采用 42℃或 40℃恒温诱导 ,rh IL- 6表达量仅为 2 0 %。

The fermentation conditions under which the human interleukin 6 (hIL 6) in recombinant E.Coli containing the temperature sensitive plasmid pKpL3a at high level on the shaker flask were investigated. The objective was to find the influence of induction strategies on the growth of recombinant cells and the expression of hIL 6. Several means of the thermal induction were used in order to enhance expression of hIL 6. The results indicated that insucha strategy of that the fermentation is carried out in 2×YT medium, 30℃, allowing the OD 600 of recombinant E.Coli up to 1.5, then the temperature is increased to 42℃ or 40℃retaining at least 1h, followed by the temperature is lowered back to 37℃ for 3h, the final expression of hIL 6, which is only 20% by single temperature upshift to 42℃ or 40℃, could be enhanced to 32%. The induction time period is less than 1h, it will not give sufficiently high expression of hIL 6. [WT5HZ]