【摘要】 目的　研究大肠杆菌重组人成骨蛋白 1的非诱导表达特性。方法　通过水质、起始pH、接种量、葡萄糖添加量、甘油添加量、装液量、转速等实验研究不同营养条件和培养条件对OP 1非诱导表达的影响。通过传代实验 ,考察该系统OP 1表达的稳定性。结果　水质对OP 1的表达没有明显影响 ,2 %～ 4 %的接种量有利于OP 1的表达 ,起始pH以 6 0为宜。葡萄糖对OP 1的表达有较大影响 ,2 %～ 4 %的葡萄糖添加量可以获得较高表达。甘油的添加能明显促进OP 1的表达 ,最适添加量为 0 3% ,较对照提高表达 81 7%。该表达系统对溶氧的需求不大 ,装液量和转速分别以 12 5～ 15 0ml和 15 0r min为宜。在优化条件下 ,最适表达时间为 2 4～ 2 6h ,OP 1的表达量可达到菌体蛋白的 4 0 %。优化的营养条件和培养条件对本系统OP 1的表达和质粒的稳定有利 ,10次传代可保持 90 %表达。结论　建立了非诱导表达的摇瓶发酵工艺 ,对其非诱导表达特性有了较为深入的了解 ,为进一步放大研究奠定了基础。更多还原

【Abstract】 Objective To study the characteristics of expression of recombinant human osteogenic prote in-1 (rhOP-1) in E.coli without induction.Methods The exp ressions of rhOP-1 under various nutrient and culture conditions,such as the quality of water,initial pH value,inoculum amount,glucose concentration,glyc erol concentration,medium volume and rotation rate,were studied.The expression s tability of the system was studied by subculture. Results The quality of water showed no significant influence on the expression of OP-1. The optimal inoculum amount and initial pH value for rhOP-1 expression were 2%-4% and 6.0 respectively.Glucose inhibited the expression of rhOP-1 significantly ,however,high expression level was obtained under a glucose concentration of 2% -4%.Glycerol accelerated the expression of rhOP-1 significantly.Under a glycer ol concentration of 0.3%,the expression level of rhOP-1 increased by 81.7% co mpared with control.The optimal medium volume and rotation rate were 125-150 ml and 150 r/min respectively.Under an optimal condition the expression level of OP-1 reached 40% of total somatic protein after a fermentation period of 24- 26 h.The optimal nutrient and culture conditions were beneficial to the expres sion of OP-1 and the stability of plasmid.After the recombinant E.coli bacteria l cells were subcultured for 10 passages,the expression of OP-1 still reached 9 0% of former level. Conclusion A fermentation procedure using spinner flask for the expression of OP-1 without induction was developed,and the characteristics of expression was studied.It la id a foundation of scale-up of production procedure of OP-1.更多还原