Cloning and functional analysis of the orotidine-5'-phosphate decarboxylase gene (PbrURA3) of the pathogenic fungus Paracoccidioides brasiliensis.
A genomic clone encoding the Paracoccidioides brasiliensis orotidine monophosphate decarboxylase gene (PbrURA3) was isolated by screening a subgenomic plasmid DNA library of this fungus, using a PCR amplification product of the gene as a probe. Sequence analysis revealed that the gene contains an open reading frame of 855 bp with a single intron (162 bp), and encodes a putative 285 amino acids polypeptide of estimated molecular weight 31.1 kDa and isoelectric point 6.5. The deduced amino acid sequence predicted a 73.4% identity with orotidine monophosphate decarboxylase of Aspergillus nidulans. Functionality of the gene was demonstrated by transformation into a Saccharomyces cerevisiae ura3 null mutant.
Reinoso
Cristina
C
Departamento de Microbiología y Genética, Universidad de Salamanca, Salamanca, Spain.
Sorais
Françoise
F
Niño-Vega
Gustavo A
GA
Fermiñán
Encarnación
E
San-Blas
Gioconda
G
Domínguez
Angel
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D015723:Gene Library; D005816:Genetic Complementation Test; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000235:genetics*; D010228:Paracoccidioides / Q000201:enzymology* / Q000235:genetics; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D014170:Transformation, Genetic
D012331:RNA, Fungal; D009964:Orotidine-5'-Phosphate Decarboxylase