Summary
Regeneration of Beauveria bassiana GK2016 protoplasts demonstrated three phases: (1) enlargement, (2) formation of a chain of budding cells, and (3) development of a true germ tube from the original protoplast. Regeneration frequencies of up to 80% were obtained when using the stabilizer ammonium sulphate. Using electroporation, protoplasts were transformed to methyl 1,2-benzimidazole carbamate (MBC) resistance with the Neurospora crassa MBC-resistant β-tubulin gene. A transformation efficiency of one of three transformants per 5 μg vector DNA was obtained. The MBC phenotype was stable and transformants grew in the presence of 5 μg MBC/ml. Southern DNA-DNA hybridization analysis demonstrated that integration of the vector into the chromosomal DNA had occurred.
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Correspondence to: G. G. Khachatourians
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Pfeifer, T.A., Khachatourians, G.G. Beauveria bassiana protoplast regeneration and transformation using electroporation. Appl Microbiol Biotechnol 38, 376–381 (1992). https://doi.org/10.1007/BF00170089
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DOI: https://doi.org/10.1007/BF00170089