Journal Article Efficient and simple electro-transformation of intact cells for the basidiomycetous fungus Pseudozyma hubeiensis

Konishi, Masaaki  ,  Yoshida, Yuta  ,  Ikarashi, Mizuki  ,  Horiuchi, Junichi  ,  小西, 正朗

8 April 2015pp.1679 - 1685 , 2015-04 , Springer Verlag
Objective: An electroporation procedure for the species was investigated to develop an efficient transformation method for the basidiomycetous fungus Pseudozyma hubeiensis SY62, a strong biosurfactant-producing host. Results: A plasmid, pUXV1emgfp including green fluorescence protein as a reporter gene, was constructed to determine the transformation and expression of foreign genes. Optimal electroporation conditions achieved 44.8 transformants μg−1 plasmid competency (intact cells) without protoplast treatment. Lithium acetate treatments increased the efficiency to approx. Twice that of control experiments. Almost all transformants demonstrated green fluorescence expressed in the transformant cells. Conclusion: The optimal method, successfully applied to several related species, yields sufficient transformant colonies to engineer the host strain.

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