Abstract:
:The macrofungus Cordyceps militaris contains many kinds of bioactive ingredients that are regulated by functional genes, but the functions of many genes in C. militaris are still unknown. In this study, to improve the frequency of homologous integration, a genetic transformation system based on a split-marker approach was developed for the first time in C. militaris to knock out a gene encoding a terpenoid synthase (Tns). The linear and split-marker deletion cassettes were constructed and introduced into C. militaris protoplasts by PEG-mediated transformation. The transformation of split-marker fragments resulted in a higher efficiency of targeted gene disruption than the transformation of linear deletion cassettes did. The color phenotype of the Tns gene deletion mutants was different from that of wild-type C. militaris. Moreover, a PEG-mediated protoplast transformation system was established, and stable genetic transformants were obtained. This method of targeted gene deletion represents an important tool for investigating the role of C. militaris genes.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Lou H,Ye Z,Yun F,Lin J,Guo L,Chen B,Mu Zdoi
10.1007/s12033-018-0080-9subject
Has Abstractpub_date
2018-05-01 00:00:00pages
380-385issue
5eissn
1073-6085issn
1559-0305pii
10.1007/s12033-018-0080-9journal_volume
60pub_type
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