重组人HBNF的表达和提纯及其生物活性的测定

目的 通过酵母表达系统表达重组人亲肝素性促轴突生长因子(heparin—binding neuritepromoting factor，HBNF)蛋白并检测其生物活性。方法 将重组表达质粒pPIC9k／HBNf通过电穿孔法转入酵母表达系统GSI15，利用G418筛选高拷贝子用于大量表达HBNF、蛋白，再检测其生物活性。结果 通过G418筛选出抗性大于2mg／mL的高拷贝子，大量表达、提纯后获得相对分子质量约18000蛋白，Western—blot证实为HBNF蛋白，生物活性检测显示提纯HBNF有显著的神经营养作用。结论 通过酵母表达系统获得的重组人HBNF有一定的生物活性。

Expression, Purification and Bio-activity Analysis of Recombined Human HBNF

Purpose To express recombined human heparin-binding neuritepromoting factor (HBNF) by yeast expression system and analyse its bio-activity. Methods Transforming the linearized pPIC9k/HBNF DNA into the competenced GS115 by electroporation,multi-copy clones were screened by G418 for scale-up expression,the bio-activity of the HBNF was analyzed. Results Multi-copy clones were screened out at a final G418 resistant concentration of 2 mg/mL,after scale-up expression,the 18 000 protein was purified,which proved to be HBNF by Western-blot,and the bio-activity analysis showed its obvious neurotrophic effects. Conclusions The rh?HBNF expressed by yeast expression system had wanted bio-activity.