Hostname: page-component-78c5997874-mlc7c Total loading time: 0 Render date: 2024-11-03T05:11:32.632Z Has data issue: false hasContentIssue false
  • English
  • Français

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
Get access

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

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)