Regulatory sequences for expressing genes in oomycete fungi.
Promoter and terminator sequences from a range of species were tested for activity in the oomycetes, a group of lower fungi that bear an uncertain taxonomic affinity to other organisms and in which little is known of the sequences required for transcription. Transient assays, using the reporter gene beta-glucuronidase (GUS), were used to examine the function of these promoters and terminators in the plant pathogens Phytophthora infestans and P. megasperma f. sp. glycinea, and in the saprophytic water mold, Achlya ambisexualis. Oomycete promoters, isolated from the ham34 and hsp70 genes of Bremia lactucae and the actin gene of P. megasperma f. sp. glycinea, resulted in high levels of GUS accumulation in each of the three oomycetes. In contrast, little or no activity was detected when promoters from higher fungi (four ascomycetes and one basidiomycete), plants, and animals were tested. The terminator from the ham34 gene resulted in much higher levels of GUS accumulation than did others, although an oomycete terminator was not absolutely required for expression. Transcript mapping of RNA from stable transformants confirmed accurate initiation from the B. lactucae hsp70 promoter and termination within 3' ham34 sequences in P. infestans. Our results indicate that the transcriptional machinery of the oomycetes differs significantly from that of the higher fungi, but that enough conservation exists within the class to allow vectors developed from one oomycete species to be used for others.
Judelson
H S
HS
NSF Center for Engineering Plants for Resistance Against Pathogens, University of California, Davis 95616.
Tyler
B M
BM
Michelmore
R W
RW
D001483:Base Sequence; D004271:DNA, Fungal; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005966:Glucuronidase / Q000235:genetics; D008969:Molecular Sequence Data; D009868:Oomycetes / Q000201:enzymology / Q000235:genetics*; D010838:Phytophthora / Q000235:genetics; D011401:Promoter Regions, Genetic; D012331:RNA, Fungal / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D013045:Species Specificity; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic; D014170:Transformation, Genetic
D004271:DNA, Fungal; D012331:RNA, Fungal; D005966:Glucuronidase