TY - JOUR
T1 - Molecular transformation of Fusarium solani with an antibiotic resistance marker having no fungal DNA homology
AU - Marek, Elzbieta T.
AU - Schardl, Christopher L.
AU - Smith, David A.
PY - 1989/6
Y1 - 1989/6
N2 - 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.[/P ]
AB - 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.[/P ]
KW - Cauliflower mosaic virus promoter
KW - Fusarium transformation
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U2 - 10.1007/BF00376799
DO - 10.1007/BF00376799
M3 - Article
C2 - 2550150
AN - SCOPUS:0024674719
SN - 0172-8083
VL - 15
SP - 421
EP - 428
JO - Current Genetics
JF - Current Genetics
IS - 6
ER -