重组酪醇糖基转移酶表达条件的优化与纯化

[目的]研究利用现代生物技术制备红景天苷的方法。[方法]试验借助SDS-PAGE方法对重组目的基因的表达条件进行了优化,比较了诱导温度、IPTG浓度、诱导时间等参数对重组基因表达的影响,以确定最佳诱导表达条件。[结果]最佳诱导表达条件为：诱导时间5 h,IPTG浓度0.6 mmol/L,诱导温度37℃;用镍柱亲和层析对该含有组氨酸标签的融合蛋白进行纯化,获得了纯度达99%以上的目的蛋白。[结论]该项研究为重组糖基转移酶的纯化提供了成功的经验。

Optimization and Purification of Expression Condition of Recombination Tyrosol Glycosyltransferase

[Objective]The aim was to study the preparation of salidroside basing on modern biotechnology.[Method]Basing on SDS-PAGE method,the expression condition of recombination gene was optimized by investigating the effects of induction temperature,concentration of IPTA,induction time on the gene expression.[Result]The optimum induction conditions were as follows∶induction time of 5 h,IPTG concentration of 0.6 mmol/L and induction temperature of 37 ℃.The target protein with purity of 99% was obtained by purifying the fusion protein with Ni-chromatography column.[Conclusion]The study provides successful experience for the purification of tyrosol glycosyltransferase.