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

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．