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用 Hind Ⅲ和 Eco RⅠ双酶酶切经改造的 P U Ca F G F质粒,获得了人酸性成纤维细胞生长因子(ha F G F)基因,将该基因片段插入表达载体pkk2233,筛选得到了重组质粒pkkha F G F。加 I P T G 诱导该质粒转化的大肠杆菌 J M 109 表达,ha F G F在发酵液中的表达水平约80m g/ L。用肝素亲和层析的方法,获得了电泳纯ha F G F样品。纯化过程中ha F G F活力回收率为65% 。纯化ha F G F样品的比活性为 E D50 4.6ng/m l,理化及生物活性分析与天然ha F G F对照品完全一致。
Human HbF gene was digested with Hind Ⅲ and EcoRI double enzyme digestion, and inserted into the expression vector pkk2233 , Screened the recombinant plasmid pkk ha F G F. Expression of this plasmid-transformed E. coli J M 109 was induced by addition of IPTG and the expression level of ha F G F in the fermentation broth was about 80 m g / L. Using heparin affinity chromatography, electrophoretic pure ha F G F samples were obtained. The ha F G F activity recovery during the purification was 65%. The specific activity of the purified ha F G F sample was E D50 4.6 ng / ml, and the physico-chemical and bioassay analyzes were in good agreement with the native ha F G F control.