纤维素降解球毛壳菌NK-102的高效遗传转化方法

Two optimized transformation protocols for a celluloseutilizing fungus Chaetomium globosum NK-102

摘要:

摘要:【目的】在球毛壳菌(Chaetomium globosum)NK-102 中，建立菌株特异性转化体系。【方法】构建新的抗性标记pUCATPH-Pgap，转化效率优于pUCATPH 和pCM768。建立了PEG-原生质体和根癌农杆菌(Agrobacterium tumefaciens)EHA105 介导的两种转化方法。【结果】原生质体转化效率为30－50 个转化子/10 μg DNA，抗性标记pUCATPH-Pgap 效率最高。EHA105 介导转化率达到3.2×102 转化子/107 孢子。Sout

Abstract:

Abstract: ［Objective］ We developed the transformation methods of the strain Chaetomium globosum NK-102．［Methods］ We constructed plasmid pUCATPH-Pgap and compared the transformation efficiency with pUCATPH and pCM768． We established the PEG mediated protoplast transformation and Agrobacterium tumefaciens EHA105 mediated transformation methods． ［Results］In protoplast approach，approximately 3－5 transformants/μg DNA could be obtained．The highest efficiency of transformation was obtained by employing pUCATPH-Pgap．A．tumefaciens EHA105 successfully mediated T-DNA insertion into the genome of C． globosum NK-102 and the transformation rate was 3.2×102 transformants /107 spores． The transformants retained stable after generations． Southern blot analyses confirmed that the DNA had integrated into the chromosomal DNA of C． globosum NK-102．［Conclusion］The transformation systems were good basis for selection of C． globosum mutant strains that effectively utilizing cellulose．