【摘要】 用产黄青霉 HY876作受体菌 ,建立 amds( Acetamidase)基因为选择标记的 VHb表达系统 .该系统构建包括原生质体的制备和再生 ;用于真菌表达 VHb基因的质粒 p VHb M和 p VHBI的构建 ;p VHb M或p VHb I与选择标记质粒 p Ucamds的共转化 ;将 VHb基因整合到产黄青霉 HY876基因组中 .结果表明 ,只有部分转化子整合有 VHb基因 ,VHb蛋白的表达量与其在染色体上整合的位点和整合拷贝数有关 ,不同转化子产生不同效应 .该菌株的摇瓶发酵实验表明 ,VHb可促进青霉素的合成 ,使青霉素效价提高 3 9% .更多还原

【Abstract】 A exogenous gene expression system was established in P. chrysogenum strain by using acetamidase(amds) gene as selection markers. The Cytoplasm-PEG transformation system was constructed including preparations and reuses of P. chrysogenum Cytoplasm and expression of exogenous gene. The two plasmids pVHbM and pVHbⅠ for expression of VHb in filamentous fungi were constructed. We were able to introduce VHb gene into P. chrysogenum by co-transformation of plasmid pVHbM or/and pVHbI with the Selection pUcamds markers. Some of transformants integrated the VHb gene. The expression level of VHb protein was related with the position and copies for integrating VHb gene into chromosome of filamentous fungi, different transformants had different efficiency. Valence of penicillin enhanced by a factor of 39% in shake-flask experiments for penicillium chrysogenun YH876 variants.更多还原