RESUMEN
<p><b>OBJECTIVE</b>To obtain the full-length cDNA of a novel gene (named yp05) associated with citrinin production-related genes in Monascus aurantiacus.</p><p><b>METHODS</b>Total RNA was extracted from mycelium, 3' and 5' cDNA end of yp05 gene was amplified using smart trace cDNA amplification kit, and the full-length cDNA of a novel gene (named yp05) was obtained from the electronic assembly of 3'-RACE and 5'-RACE products.</p><p><b>RESULTS</b>This yp05 gene was 787 bp including a 597 bp open reading frame (ORF) and encoded a deduced protein with 199 amino acid residues, and the amino acid sequence of this protein was found similar with the sequences of many fungal manganese-superoxide dismutases in the GenBank with the aid of BLASTp. The transcription of yp05 gene in Monascus strains was analyzed with the aid of Northern blotting. The transcription of yp05 gene was only detected in Monascus strains, provided that citrinin was produced.</p><p><b>CONCLUSION</b>The transcription of yp05 gene belongs to differential expression genes of citrinin yielded from Monascus and has no correlation with the biosynthesis pathway of red pigments.</p>
Asunto(s)
Secuencia de Aminoácidos , Secuencia de Bases , Northern Blotting , Citrinina , Clonación Molecular , ADN Complementario , Química , Proteínas Fúngicas , Química , Genética , Biblioteca de Genes , Datos de Secuencia Molecular , Monascus , Genética , Metabolismo , Micelio , Genética , Metabolismo , Pigmentos Biológicos , ARN Mensajero , Metabolismo , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
<p><b>OBJECTIVE</b>To construct a tag expression library of Monascus aurantiacus that could produce citrinin maximally on the thirteenth (0.966 mg/mL) day in the submerged culture.</p><p><b>METHODS</b>Total RNA was extracted from the mycelium, cDNA was synthesized using the SuperScript choice system, and then, a SAGE library was successfully constructed according to the MicroSAGE method.</p><p><b>RESULTS</b>Five hundred and ninety eight clones were obtained in SAGE library, and 120 clones were picked out randomly for identification and sequencing purpose. Eighty nine clones had positive inserts, 26 clones had no inserts and the remaining 5 clones had no site of NlaIII enzyme in inserts. There were seven repeated clones.</p><p><b>CONCLUSION</b>With the aid of SAGE2000 software, 901 tags were obtained from 89 clones, representing 686 unique transcripts. Six unique tags of them belong to highly expressed genes (Number of tags > or = 10) and 143 unique tags to moderately expressed genes (repeat tags > or = 2).</p>