Mitochondria1 DNA (mt DNA) from a patulin producer, Penicillium urticue (synonym P. griseofulvum), was 27.8kb+0.6kb in size by electron microscopy and 27.2kb by agarose gel electrophoresis. Restriction endonuclease maps for nine restriction enzymes were constructed, and eleven fragments which. covered the total range of themt DNA were cloned into the Escherichk coli plasmid vector pUCl9. Southern analysis of the native genomes of P. urtkue amlE chrysogenum with six of the cloned fragments as probes indicated similar genome arrangements as well as similar restriction mps.=.Both.the large and small rRNA genes of P. urticae and P. chrysogenum were located on these restriction maps using Southern hybridization,.and the result also supported the similar arrangement. Agaroselformaldehyde gel electrophoresis indicated that the small rRNA was 1.5 kb in size in both species; but, surprisingly, the large rRNA was 4.2 kb in size for P. d a e and 3.5 kb for P. chrysogenum.These sizes were, respectively, 1-1 kb and 0-4 kb larger than those from the very closely related Aspergillus nirlulans.
The nucleotide sequence of a large rRNA gene and its flanking regions in cloned fragments of mitochondrial DNA from a patulin producer, Penicillium urticae NRRL2159A, was determined by dideoxy sequencing, and the 5' end and intron-exon border of the 1-rRNA gene were determined by primer extension analysis and RNA sequencing, respectively. In addition to the extensive sequence homology of the 3' end of the P. urticae mt 1-rRNA gene with those of Aspergillus nidulans and Neurospora crassa, the P. urticae gene had a 1,685 bp intron which separates a 3,307 bp 5' exon and a 583 bp 3' exon. In spite of being closely related Penicillium species, the size of the 5' exon of the P. urticae mt 1-rRNA is 472 bp larger than that of P. chrysogenum, whereas the sizes of the 3' exon and intron of P. urticae are very similar to those of P. chrysogenum (581 bp for the 3' exon and 1,678 bp for the intron). The intron of P. urticae contains a structure similar to the consensus one of the self splicing group IA intron and a large open reading frame suggested to be a gene for ribosomal protein S5. A sequence similar to the I-SceI recognition sequence was found at the exon-intron border. Extensive sequence homology was observed between P. urticae and P. chrysogenum, exceptions being in four regions in the 5' exon. These non-homologous regions were located in the hairpin and variable regions outside of the core structures. Comparison of the mt 1-rRNA sequences of several filamentous fungi revealed that the above four non-homologous regions are greatly expanded, and two other non-homologous regions appear at the 3' ends of the 5' exon and 3' exon.
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