Co-transformation of Metarhizium anisopliae by electroporation or using the gene gun to produce stable GUS transformants
FEMS Microbiology Letters
The potential of β-glucuronidase as a molecular marker for studying the environmental microbiology of entomopathogenic fungi was assessed. Metarhizium anisopliae was stably co-transformed with plasmids (pNOM102 and pBENA3) containing the β-glucuronidase and benomyl resistance ( β-tubulin) genes, using both electroporation and biolistic delivery systems, and it was confirmed that the expressed phenotypes were not exhibited by ten randomly chosen indigenous North-American isolates. In spite of random and multiple integrations, the co-transformants showed normal growth rates and retained their pathogenicity to insects. β-Glucuronidase activity in the co-transformants was used to detect histochemically the presence of fungal hyphae in infected host insects (Bombyx mori) and thus provides a practical means of marking genetically engineered pathogens for field trials.
St. Leger, R.J., S. Shimizu, M.J. Bidochka and D.W. Roberts. 1995. Co-transformation of Metarhizium anisopliae by electroporation or using the gene gun to produce stable GUS transformants. FEMS Microbiol Letters 131: 289-294.