毕赤酵母高效表达血小板源生长因子研究

Studies on the High Effective Expression of Pletelet-derived Growth Factor(P DGF) in the Pichia pastoris Yeast

血小板源生长因子(PDGF)是由多种细胞产生的肽类生长因子,在细胞培养、皮肤溃疡的治疗以及化妆品添加剂中有很重要的作用.将编码PDGF基因克隆到表达载体Xl-1-Blue上,通过电转化整合到毕赤酵母的基因组中,在毕赤酵母三磷酸甘油醛脱氢酶(GAP)启动子的作用下,表达PDGF蛋白,其分子质量为30kD左右,估计其表达量为80～100mg/L.采用MTT法也证实其生物学活性与天然PDGF蛋白非常相似.

Pletelet-derived growth factor, peptide growth factor, plays an important role in cell cultivation and the treatment of skin ulceration and is an important additive of cosmetic. PDGF gene was cloned into the Pichia Pastoris expression vector X1-1-Blue, then transformed into chromosome of Pichia Pastoris SMD1168 strain by electroporation and expressed by the action of promoter GAP. The expressed PDGF protein has a molecular weight of about 30 kD and its expression capacity is from 80 - 100 mg/L. Activity measurement of PDGF protein produced from Pichia Pastoris Yeast by the was similar to the natural PDGF protein. method of MTT showed that its biological activity