高等学校化学学报

高等学校化学学报 ›› 2004, Vol. 25 ›› Issue (5): 858.

• 研究论文 • 上一篇    下一篇

产黄青霉菌中细菌血红蛋白的基因表达及工程菌的应用研究

朱彦民, 刘晓均, 孙妍红, 张今   

  1. 吉林大学分子酶学工程教育部重点实验室, 长春 130023
  • 收稿日期:2003-05-09 出版日期:2004-05-24 发布日期:2004-05-24
  • 通讯作者: 张　今(1940年出生),男,教授,博士生导师,从事基因工程和蛋白质工程研究.E-mail:zhangjin@mai.ljlu.edu.cn E-mail:zhangjin@mai.ljlu.edu.cn

Expression and Application of a Baterial Hemoglobin Gene in Penicillium Chrysogenum

ZHU Yan-Min, LIU Xiao-Jun, SUN Yan-Hong, ZHANG Jin   

  1. Key Lab for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun 130023, China
  • Received:2003-05-09 Online:2004-05-24 Published:2004-05-24

PDF (PC)

被引次数

6

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pVHbM和pVHBI的构建;pVHbM或pVHbI与选择标记质粒pUcamds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39%.

关键词: 细菌血红蛋白基因, 表达, 产黄青霉素菌

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.

Key words: Vitreosicilla hemoglobin gene, Expression, Chrysogenum

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