In vivo addition of telomeric repeats to foreign DNA generates extrachromosomal DNAs in the taxol-producing fungus Pestalotiopsis microspora.
Transformation of the taxol-producing filamentous fungus Pestalotiopsis microspora with a plasmid containing the bacterial hygromycin resistance gene fused to Aspergillus regulatory sequences resulted in the in vivo formation of extrachromosomal DNAs with telomeric repeats in the majority of transformants. Repeats of the telomeric sequence 5'-TTAGGG-3' were appended to nontelomeric transforming DNA termini. No fungal sequences other than telomeric repeats were detected in extrachromosomal DNAs. Transformants contained three to six different sizes or conformational forms of extrachromosomal DNAs. The DNAs showed no change in size or internal structure during 6 months of growth with selection, but were lost after 20 days of growth without selection. Transformation of wild-type P. microspora with a PCR-amplified extrachromosomal DNA having terminal telomeric repeats produced up to 50-fold more transformants than the original transformation vector. The addition of telomeric repeats to foreign DNA is unusual among fungi and may have important adaptive or developmental implications.
Department of Plant Sciences, Montana State University, Bozeman, Montana 59717, USA. long@montana.edu
D M
DM
Long
E D
ED
Smidansky
A J
AJ
Archer
G A
GA
Strobel
D001233:Aspergillus nidulans / Q000235:genetics; D001234:Aspergillus niger / Q000235:genetics; D001483:Base Sequence; D004352:Drug Resistance, Microbial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005800:Genes, Fungal; D005821:Genetic Techniques; D003904:Mitosporic Fungi / Q000235:genetics* / Q000378:metabolism; D017239:Paclitaxel / Q000096:biosynthesis*; D010641:Phenotype; D010957:Plasmids; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D016615:Telomere / Q000235:genetics*; D014162:Transfection; D014170:Transformation, Genetic; D014197:Trees / Q000382:microbiology
D017239:Paclitaxel