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Transformation of sapstaining fungi with hygromycin B resistance plasmids pAN7-1 and pCB1004

Published online by Cambridge University Press:  01 January 1999

HONG-LONG WANG ,
SEONG HWAN KIM ,
HENRY SIU  and
COLETTE BREUIL
HONG-LONG WANG
Affiliation:
Chair of Forest Products Biotechnology, Department of Wood Science, University of British Columbia, Vancouver, Canada V6T 1Z4
SEONG HWAN KIM
Affiliation:
Chair of Forest Products Biotechnology, Department of Wood Science, University of British Columbia, Vancouver, Canada V6T 1Z4
HENRY SIU
Affiliation:
Chair of Forest Products Biotechnology, Department of Wood Science, University of British Columbia, Vancouver, Canada V6T 1Z4
COLETTE BREUIL
Affiliation:
Chair of Forest Products Biotechnology, Department of Wood Science, University of British Columbia, Vancouver, Canada V6T 1Z4
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Abstract

Protoplasts were produced by treating the yeast-like cells of O. piceae and O. quercus with high concentrations of Novozym 234 (20 mg ml−1). Transformants with resistance to hygromycin B were obtained by transforming the protoplasts with the plasmids pAN7-1 and pCB1004 containing the hygromycin B phosphotransferase (hph) gene. For all isolates tested, transformation efficiencies ranged between 103 and 105 transformants μg−1 vector DNA. High polyethylene glycol concentration (66 %) increased the transformation efficiency. Integration of the hph gene into the transformant genome was verified by a polymerase chain reaction and dot DNA hybridization analysis. There was no evidence of mitotic instability of hygromycin B resistance in the transformants.

Type
Research Article
Information
Mycological Research , Volume 103 , Issue 1 , January 1999 , pp. 77 - 80
Copyright
© The British Mycological Society 1999

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