重组拖丝蛋白基因在大肠杆菌中表达条件的优化

Expression Conditions of Recombinant Spider Dragline Silk Protein in Escherichia coli.

对已构建的重组蜘蛛拖丝蛋白基因表达菌株 p NS2 ,以 IPTG为诱导剂 ,建立最适表达条件 .在 LB培养基中添加质量分数为 0 .1 %的甘氨酸和丙氨酸 ,菌体培养至密度 A6 0 0 =0 .5～ 0 .7时加入浓度为 0 .2 mmol/ L的 IPTG,诱导 5 h,可得到表达量占可溶性总蛋白质 2 7.2 %的较好结果 .

We studied on some factors affecting the expression of the recombinant spider dragline silk protein production in Escherichia coli.. The results showed that the optimal induce conditions for expression of recombinant spider dragline silk protein are follows: add 0.1% glycine and 0.1% lactamine in LB medium, expression of the spider silks protein was induced by the supply of IPTG to a final concentration of 0.2 mmol/L when cultures were grown to A 600=0.5～0.7, and the fusion gene highly expressed 5 h later. The expressing product is 27.2% of the total bacterial protein.