摘要
【目的】山蛭素基因在甲醇毕赤酵母中表达。【方法】利用PCR点突变方法改造山蛭素基因 ,将山蛭素基因克隆到pGEM Teasy载体中。对重组质粒测序 ,构建了毕赤酵母表达载体 pPIZB Hs ,然后转化有活性的GS115酵母菌株 ,筛选表达酵母菌株GS115 ,摇瓶发酵。【结果】改造的山蛭素基因在重组酵母中表达 ,重组蛋白质具有对凝血酶抑制活性。【结论】利用PCR点突变方法能改造山蛭素基因 。
To study the expression of Hs gene in methylotrophic yeast pichia pastoris. Haemadin gene(from Haemadipsa sylvestris, Hs gene)was modified with PCR site directed mutagenesis methods. Hs gene was cloned into pGEM T easy vector. The recombinant plasmid was sequenced. The expression vector pPICZB Hs was constructed. Competent GS115 yeast strain was transformed by chemical transformation method based on the use of PEG. The expression GS115 yeast strain was selected. Flask fermentation of recombinant yeast was carried out. Hs gene was successfully modified. Modified Hs gene was expressed in recombinant yeast. The recombinant protein had thrombin inhibitory activity. [Conclusion] Hs gene can be modified with PCR site directed mutagenesis methods,and can express in yeast pichia pastoris.
出处
《中山医科大学学报》
CSCD
北大核心
2002年第1期21-23,共3页
Academic Journal of Sun Yat-sen University of Medical Sciences
基金
国家自然科学基金资助项目 ( 3880 0 6 8.3916 0 0 17)
