Premium
Notes High‐efficiency transformation system for the biocontrol agents, Trichoderma spp.
Author(s) -
HerreraEstrella A.,
Goldman G. H.,
Montagu M. Van
Publication year - 1990
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1990.tb00654.x
Subject(s) - biology , selectable marker , transformation (genetics) , plasmid , aspergillus nidulans , hygromycin b , southern blot , trichoderma , homologous recombination , heterologous , replicon , dna , microbiology and biotechnology , genetics , gene , botany , mutant
Summary We have developed an efficient transformation system based on the use of polyethylene glycol and CaCl 2 for the biocontrol agents, Trichoderma spp. Trans formation was obtained with the plasmid pAN7‐1, carrying a bacterial hygromycin‐resistance gene as a selectable marker, under the control of Aspergillus nidulans heterologous expression signals. The system described here yielded 200‐800 transformants per μg of DNA. Transformants contained several copies of the plasmid integrated into their genome, apparently at the same site in the different transformants analysed. Stability of the transformants was achieved by inserting a 2.4kb homologous DNA fragment into pAN7‐1. Southern blot analysis indicated that integration in the stable transformants occurs through non‐homologous recombination.