Molecular transformation of Fusarium solani with an antibiotic resistance marker having no fungal DNA homology.

Abstract:

:A vector was constructed for transformation of the plant pathogenic fungus Fusarium solani. The promoter 35Sp, from cauliflower mosaic virus, was fused to the bacterial gene APH(3')II, which confers resistance to the aminoglycoside antibiotic G418. Two transformation procedures were developed: one using isolated fungal protoplasts, the other using germinated fungal spores. A transformation frequency of 3.3 G418-resistant colonies were obtained per microgram DNA. Of 14 colonies analyzed, 12 had vector sequences integrated into their high molecular weight DNA, and 2 were untransformed. Integration was sometimes accompanied by rearrangements of both the vector and flanking fungal DNAs. Primer-extension analysis of the mRNA from one transformant revealed two putative transcription initiation sites in the chimeric APH(3')II gene. Both sites differed from the normal initiation site in plants. This vector will be useful in transformation systems in which integration by non-homologous recombination is desired.

journal_name

Curr Genet

journal_title

Current genetics

authors

Marek ET,Schardl CL,Smith DA

doi

10.1007/BF00376799

subject

Has Abstract

pub_date

1989-06-01 00:00:00

pages

421-8

issue

6

eissn

0172-8083

issn

1432-0983

journal_volume

15

pub_type

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