Co-transformation of Metarhizium Anisopliae by Electroporation or Using the Gene Gun to Produce Stable GUS Transformants

Document Type

Article

Journal/Book Title/Conference

FEMS Microbiology Letters

Volume

131

Issue

3

Publication Date

9-15-1995

First Page

289

Last Page

294

Abstract

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.

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